Clathrin: The main structural coat protein of COATED VESICLES which play a key role in the intracellular transport between membranous organelles. Each molecule of clathrin consists of three light chains (CLATHRIN LIGHT CHAINS) and three heavy chains (CLATHRIN HEAVY CHAINS) that form a structure called a triskelion. Clathrin also interacts with cytoskeletal proteins.Clathrin Heavy Chains: The heavy chain subunits of clathrin.Clathrin Light Chains: The light chain subunits of clathrin.Monomeric Clathrin Assembly Proteins: A subclass of clathrin assembly proteins that occur as monomers.Clathrin-Coated Vesicles: Vesicles formed when cell-membrane coated pits (COATED PITS, CELL-MEMBRANE) invaginate and pinch off. The outer surface of these vesicles is covered with a lattice-like network of the protein CLATHRIN. Shortly after formation, however, the clathrin coat is removed and the vesicles are referred to as ENDOSOMES.Coated Pits, Cell-Membrane: Specialized regions of the cell membrane composed of pits coated with a bristle covering made of the protein CLATHRIN. These pits are the entry route for macromolecules bound by cell surface receptors. The pits are then internalized into the cytoplasm to form the COATED VESICLES.Adaptor Protein Complex 2: An adaptor protein complex primarily involved in the formation of clathrin-related endocytotic vesicles (ENDOSOMES) at the CELL MEMBRANE.Adaptor Proteins, Vesicular Transport: A class of proteins involved in the transport of molecules via TRANSPORT VESICLES. They perform functions such as binding to the cell membrane, capturing cargo molecules and promoting the assembly of CLATHRIN. The majority of adaptor proteins exist as multi-subunit complexes, however monomeric varieties have also been found.Endocytosis: Cellular uptake of extracellular materials within membrane-limited vacuoles or microvesicles. ENDOSOMES play a central role in endocytosis.Adaptor Protein Complex alpha Subunits: A family of large adaptin protein subunits of approximately 100 kDa in size. They have been primarily found as components of ADAPTOR PROTEIN COMPLEX 2.Adaptor Protein Complex 1: A clathrin adaptor protein complex primarily involved in clathrin-related transport at the TRANS-GOLGI NETWORK.Coated Vesicles: Vesicles formed when cell-membrane coated pits (COATED PITS, CELL-MEMBRANE) invaginate and pinch off. The outer surface of these vesicles are covered with a lattice-like network of coat proteins, such as CLATHRIN, coat protein complex proteins, or CAVEOLINS.Auxilins: A family of proteins that play a role as cofactors in the process of CLATHRIN recycling in cells.Endosomes: Cytoplasmic vesicles formed when COATED VESICLES shed their CLATHRIN coat. Endosomes internalize macromolecules bound by receptors on the cell surface.trans-Golgi Network: A network of membrane compartments, located at the cytoplasmic side of the GOLGI APPARATUS, where proteins and lipids are sorted for transport to various locations in the cell or cell membrane.Adaptor Protein Complex beta Subunits: A family of large adaptin protein complex subunits of approximately 90-130 kDa in size.HSC70 Heat-Shock Proteins: A constitutively expressed subfamily of the HSP70 heat-shock proteins. They preferentially bind and release hydrophobic peptides by an ATP-dependent process and are involved in post-translational PROTEIN TRANSLOCATION.Dynamins: A family of high molecular weight GTP phosphohydrolases that play a direct role in vesicle transport. They associate with microtubule bundles (MICROTUBULES) and are believed to produce mechanical force via a process linked to GTP hydrolysis. This enzyme was formerly listed as EC 3.6.1.50.Adaptor Protein Complex mu Subunits: A family of medium adaptin protein subunits of approximately 45 KDa in size. They have been primarily found as components of ADAPTOR PROTEIN COMPLEX 3 and ADAPTOR PROTEIN COMPLEX 4.Adaptor Protein Complex gamma Subunits: A family of large adaptin protein subunits of approximately 90 KDa in size. They have been primarily found as components of ADAPTOR PROTEIN COMPLEX 1.Adaptor Protein Complex 3: An adaptor protein complex found primarily on perinuclear compartments.Protein Transport: The process of moving proteins from one cellular compartment (including extracellular) to another by various sorting and transport mechanisms such as gated transport, protein translocation, and vesicular transport.Vesicular Transport Proteins: A broad category of proteins involved in the formation, transport and dissolution of TRANSPORT VESICLES. They play a role in the intracellular transport of molecules contained within membrane vesicles. Vesicular transport proteins are distinguished from MEMBRANE TRANSPORT PROTEINS, which move molecules across membranes, by the mode in which the molecules are transported.Protein Binding: The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments.Transferrin: An iron-binding beta1-globulin that is synthesized in the LIVER and secreted into the blood. It plays a central role in the transport of IRON throughout the circulation. A variety of transferrin isoforms exist in humans, including some that are considered markers for specific disease states.Cattle: Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.Golgi Apparatus: A stack of flattened vesicles that functions in posttranslational processing and sorting of proteins, receiving them from the rough ENDOPLASMIC RETICULUM and directing them to secretory vesicles, LYSOSOMES, or the CELL MEMBRANE. The movement of proteins takes place by transfer vesicles that bud off from the rough endoplasmic reticulum or Golgi apparatus and fuse with the Golgi, lysosomes or cell membrane. (From Glick, Glossary of Biochemistry and Molecular Biology, 1990)ADP-Ribosylation Factors: MONOMERIC GTP-BINDING PROTEINS that were initially recognized as allosteric activators of the MONO(ADP-RIBOSE) TRANSFERASE of the CHOLERA TOXIN catalytic subunit. They are involved in vesicle trafficking and activation of PHOSPHOLIPASE D. This enzyme was formerly listed as EC 3.6.1.47Membrane Proteins: Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors.Freeze Etching: A replica technique in which cells are frozen to a very low temperature and cracked with a knife blade to expose the interior surfaces of the cells or cell membranes. The cracked cell surfaces are then freeze-dried to expose their constituents. The surfaces are now ready for shadowing to be viewed using an electron microscope. This method differs from freeze-fracturing in that no cryoprotectant is used and, thus, allows for the sublimation of water during the freeze-drying process to etch the surfaces.Cell Membrane: The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.Dynamin I: A subtype of dynamin found primarily in the NEURONS of the brain.Molecular Sequence Data: Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.Arrestins: Regulatory proteins that down-regulate phosphorylated G-protein membrane receptors, including rod and cone photoreceptors and adrenergic receptors.Microscopy, Fluorescence: Microscopy of specimens stained with fluorescent dye (usually fluorescein isothiocyanate) or of naturally fluorescent materials, which emit light when exposed to ultraviolet or blue light. Immunofluorescence microscopy utilizes antibodies that are labeled with fluorescent dye.Amino Acid Sequence: The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.HeLa Cells: The first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for VIRUS CULTIVATION and antitumor drug screening assays.Transport Vesicles: Vesicles that are involved in shuttling cargo from the interior of the cell to the cell surface, from the cell surface to the interior, across the cell or around the cell to various locations.Nerve Tissue ProteinsPhosphoproteinsRecombinant Fusion Proteins: Recombinant proteins produced by the GENETIC TRANSLATION of fused genes formed by the combination of NUCLEIC ACID REGULATORY SEQUENCES of one or more genes with the protein coding sequences of one or more genes.Adaptor Protein Complex sigma Subunits: A family of small adaptin protein complex subunits of approximately 19 KDa in size.Carrier Proteins: Transport proteins that carry specific substances in the blood or across cell membranes.Adaptor Protein Complex delta Subunits: A family of large adaptin protein subunits of approximately 130-kDa in size. They have been primarily found as components of ADAPTOR PROTEIN COMPLEX 3.Microscopy, Electron: Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen.Amino Acid Motifs: Commonly observed structural components of proteins formed by simple combinations of adjacent secondary structures. A commonly observed structure may be composed of a CONSERVED SEQUENCE which can be represented by a CONSENSUS SEQUENCE.Transcription Factor AP-2: A family of DNA binding proteins that regulate expression of a variety of GENES during CELL DIFFERENTIATION and APOPTOSIS. Family members contain a highly conserved carboxy-terminal basic HELIX-TURN-HELIX MOTIF involved in dimerization and sequence-specific DNA binding.Macromolecular Substances: Compounds and molecular complexes that consist of very large numbers of atoms and are generally over 500 kDa in size. In biological systems macromolecular substances usually can be visualized using ELECTRON MICROSCOPY and are distinguished from ORGANELLES by the lack of a membrane structure.HSP70 Heat-Shock Proteins: A class of MOLECULAR CHAPERONES found in both prokaryotes and in several compartments of eukaryotic cells. These proteins can interact with polypeptides during a variety of assembly processes in such a way as to prevent the formation of nonfunctional structures.Protein Structure, Tertiary: The level of protein structure in which combinations of secondary protein structures (alpha helices, beta sheets, loop regions, and motifs) pack together to form folded shapes called domains. Disulfide bridges between cysteines in two different parts of the polypeptide chain along with other interactions between the chains play a role in the formation and stabilization of tertiary structure. Small proteins usually consist of only one domain but larger proteins may contain a number of domains connected by segments of polypeptide chain which lack regular secondary structure.Caveolae: Endocytic/exocytic CELL MEMBRANE STRUCTURES rich in glycosphingolipids, cholesterol, and lipid-anchored membrane proteins that function in ENDOCYTOSIS (potocytosis), transcytosis, and SIGNAL TRANSDUCTION. Caveolae assume various shapes from open pits to closed vesicles. Caveolar coats are composed of CAVEOLINS.Cell Line: Established cell cultures that have the potential to propagate indefinitely.ADP-Ribosylation Factor 1: ADP-RIBOSYLATION FACTOR 1 is involved in regulating intracellular transport by modulating the interaction of coat proteins with organelle membranes in the early secretory pathway. It is a component of COAT PROTEIN COMPLEX I. This enzyme was formerly listed as EC 3.6.1.47.Binding Sites: The parts of a macromolecule that directly participate in its specific combination with another molecule.Green Fluorescent Proteins: Protein analogs and derivatives of the Aequorea victoria green fluorescent protein that emit light (FLUORESCENCE) when excited with ULTRAVIOLET RAYS. They are used in REPORTER GENES in doing GENETIC TECHNIQUES. Numerous mutants have been made to emit other colors or be sensitive to pH.COS Cells: CELL LINES derived from the CV-1 cell line by transformation with a replication origin defective mutant of SV40 VIRUS, which codes for wild type large T antigen (ANTIGENS, POLYOMAVIRUS TRANSFORMING). They are used for transfection and cloning. (The CV-1 cell line was derived from the kidney of an adult male African green monkey (CERCOPITHECUS AETHIOPS).)

Arrestin function in G protein-coupled receptor endocytosis requires phosphoinositide binding. (1/1930)

Internalization of agonist-activated G protein-coupled receptors is mediated by non-visual arrestins, which also bind to clathrin and are therefore thought to act as adaptors in the endocytosis process. Phosphoinositides have been implicated in the regulation of intracellular receptor trafficking, and are known to bind to other coat components including AP-2, AP180 and COPI coatomer. Given these observations, we explored the possibility that phosphoinositides play a role in arrestin's function as an adaptor. High-affinity binding sites for phosphoinositides in beta-arrestin (arrestin2) and arrestin3 (beta-arrestin2) were identified, and dissimilar effects of phosphoinositide and inositol phosphate on arrestin interactions with clathrin and receptor were characterized. Alteration of three basic residues in arrestin3 abolished phosphoinositide binding with complete retention of clathrin and receptor binding. Unlike native protein, upon agonist activation, this mutant arrestin3 expressed in COS1 cells neither supported beta2-adrenergic receptor internalization nor did it concentrate in coated pits, although it was recruited to the plasma membrane. These findings indicate that phosphoinositide binding plays a critical regulatory role in delivery of the receptor-arrestin complex to coated pits, perhaps by providing, with activated receptor, a multi-point attachment of arrestin to the plasma membrane.  (+info)

Human granulocytic ehrlichiosis agent and Ehrlichia chaffeensis reside in different cytoplasmic compartments in HL-60 cells. (2/1930)

The human granulocytic ehrlichiosis (HGE) agent resides and multiplies exclusively in cytoplasmic vacuoles of granulocytes. Double immunofluorescence labeling was used to characterize the nature of the HGE agent replicative inclusions and to compare them with inclusions containing the human monocytic ehrlichia, Ehrlichia chaffeensis, in HL-60 cells. Although both Ehrlichia spp. can coinfect HL-60 cells, they resided in separate inclusions. Inclusions of both Ehrlichia spp. were not labeled with either anti-lysosome-associated membrane protein 1 or anti-CD63. Accumulation of myeloperoxidase-positive granules were seen around HGE agent inclusions but not around E. chaffeensis inclusions. 3-(2, 4-Dinitroanilino)-3'-amino-N-methyldipropylamine and acridine orange were not localized to either inclusion type. Vacuolar-type H+-ATPase was not colocalized with HGE agent inclusions but was weakly colocalized with E. chaffeensis inclusions. E. chaffeensis inclusions were labeled with the transferrin receptor, early endosomal antigen 1, and rab5, but HGE agent inclusions were not. Some HGE agent and E. chaffeensis inclusions colocalized with major histocompatibility complex class I and II antigens. These two inclusions were not labeled for annexins I, II, IV, and VI; alpha-adaptin; clathrin heavy chain; or beta-coatomer protein. Vesicle-associated membrane protein 2 colocalized to both inclusions. The cation-independent mannose 6-phosphate receptor was not colocalized with either inclusion type. Endogenously synthesized sphingomyelin, from C6-NBD-ceramide, was not incorporated into either inclusion type. Brefeldin A did not affect the growth of either Ehrlichia sp. in HL-60 cells. These results suggest that the HGE agent resides in inclusions which are neither early nor late endosomes and does not fuse with lysosomes or Golgi-derived vesicles, while E. chaffeensis resides in an early endosomal compartment which accumulates the transferrin receptor.  (+info)

Identification of a novel domain shared by putative components of the endocytic and cytoskeletal machinery. (3/1930)

We have identified a approximately 140 amino acid domain that is shared by a variety of proteins in budding and fission yeast, nematode, rat, mouse, frog, oat, and man. Typically, this domain is located within 20 residues of the N-terminus of the various proteins. The percent identity among the domains in the 12 proteins ranges from 42 to 93%, with 16 absolutely conserved residues: N-x(11-13)-V-x2-A-T-x(34-36)-R-x(7-8)-W-R-x3-K-x12-G-x-E-x15 -L-x11-12-D-x-G-R-x11-D-x7-R. Even though these proteins share little beyond their segment of homology, data are emerging that several of the proteins are involved in endocytosis and or regulation of cytoskeletal organization. We have named this protein segment the ENTH domain, for Epsin N-terminal Homology domain, and hypothesize that it is a candidate for binding specific ligands and/or enzymatic activity in the cell.  (+info)

The EH and SH3 domain Ese proteins regulate endocytosis by linking to dynamin and Eps15. (4/1930)

Clathrin-mediated endocytosis is a multistep process which requires interaction between a number of conserved proteins. We have cloned two mammalian genes which code for a number of endocytic adaptor proteins. Two of these proteins, termed Ese1 and Ese2, contain two N-terminal EH domains, a central coiled-coil domain and five C-terminal SH3 domains. Ese1 is constitutively associated with Eps15 proteins to form a complex with at least 14 protein-protein interaction surfaces. Yeast two-hybrid assays have revealed that Ese1 EH and SH3 domains bind epsin family proteins and dynamin, respectively. Overexpression of Ese1 is sufficient to block clathrin-mediated endocytosis in cultured cells, presumably through disruption of higher order protein complexes, which are assembled on the endogenous Ese1-Eps15 scaffold. The Ese1-Eps15 scaffold therefore links dynamin, epsin and other endocytic pathway components.  (+info)

AP-4, a novel protein complex related to clathrin adaptors. (5/1930)

Here we report the identification and characterization of AP-4, a novel protein complex related to the heterotetrameric AP-1, AP-2, and AP-3 adaptors that mediate protein sorting in the endocytic and late secretory pathways. The key to the identification of this complex was the cloning and sequencing of two widely expressed, mammalian cDNAs encoding new homologs of the adaptor beta and sigma subunits named beta4 and sigma4, respectively. An antibody to beta4 recognized in human cells an approximately 83-kDa polypeptide that exists in both soluble and membrane-associated forms. Gel filtration, sedimentation velocity, and immunoprecipitation experiments revealed that beta4 is a component of a multisubunit complex (AP-4) that also contains the sigma4 polypeptide and two additional adaptor subunit homologs named mu4 (mu-ARP2) and epsilon. Immunofluorescence analyses showed that AP-4 is associated with the trans-Golgi network or an adjacent structure and that this association is sensitive to the drug brefeldin A. We propose that, like the related AP-1, AP-2, and AP-3 complexes, AP-4 plays a role in signal-mediated trafficking of integral membrane proteins in mammalian cells.  (+info)

A modulatory role for clathrin light chain phosphorylation in Golgi membrane protein localization during vegetative growth and during the mating response of Saccharomyces cerevisiae. (6/1930)

The role of clathrin light chain phosphorylation in regulating clathrin function has been examined in Saccharomyces cerevisiae. The phosphorylation state of yeast clathrin light chain (Clc1p) in vivo was monitored by [32P]phosphate labeling and immunoprecipitation. Clc1p was phosphorylated in growing cells and also hyperphosphorylated upon activation of the mating response signal transduction pathway. Mating pheromone-stimulated hyperphosphorylation of Clc1p was dependent on the mating response signal transduction pathway MAP kinase Fus3p. Both basal and stimulated phosphorylation occurred exclusively on serines. Mutagenesis of Clc1p was used to map major phosphorylation sites to serines 52 and 112, but conversion of all 14 serines in Clc1p to alanines [S(all)A] was necessary to eliminate phosphorylation. Cells expressing the S(all)A mutant Clc1p displayed no defects in Clc1p binding to clathrin heavy chain, clathrin trimer stability, sorting of a soluble vacuolar protein, or receptor-mediated endocytosis of mating pheromone. However, the trans-Golgi network membrane protein Kex2p was not optimally localized in mutant cells. Furthermore, pheromone treatment exacerbated the Kex2p localization defect and caused a corresponding defect in Kex2p-mediated maturation of the alpha-factor precursor. The results reveal a novel requirement for clathrin during the mating response and suggest that phosphorylation of the light chain subunit modulates the activity of clathrin at the trans-Golgi network.  (+info)

Enhancement of endocytosis due to aminophospholipid transport across the plasma membrane of living cells. (7/1930)

Formation of intracellular vesicles is initiated by membrane budding. Here we test the hypothesis that the plasma membrane surface area asymmetry could be a driving force for vesicle formation during endocytosis. The inner layer phospholipid number was therefore increased by adding exogenous aminophospholipids to living cells, which were then translocated from the outer to the inner layer of the membrane by the ubiquitous flippase. Addition of either phosphatidylserine or phosphatidylethanolamine led to an enhancement of endocytosis, showing that the observed acceleration does not depend on the lipid polar head group. Conversely, a closely related aminophospholipid that is not recognized by the flippase, lyso-alpha-phosphatidylserine, inhibited endocytosis, and similar results were obtained with a cholesterol derivative that also remains in the plasma membrane outer layer. Thus an increase of lipid concentration in the inner layer enhanced internalization, whereas an increase of the lipid concentration in the outer layer inhibited internalization. These experiments suggest that transient asymmetries in lipid concentration might contribute to the formation of endocytic vesicles.  (+info)

Entry of porcine reproductive and respiratory syndrome virus into porcine alveolar macrophages via receptor-mediated endocytosis. (8/1930)

Porcine alveolar macrophages (AMphi) are the dominant cell type that supports the replication of porcine reproductive and respiratory syndrome virus (PRRSV) in vivo and in vitro. In order to determine the characteristics of the virus-receptor interaction, the attachment of PRRSV to cells was examined by using biotinylated virus in a series of flow cytometric assays. PRRSV bound specifically to AMphi in a dose-dependent manner. Binding of PRRSV to AMphi increased gradually and reached a maximum within 60 min at 4 degrees C. By confocal microscopy, it was shown that different degrees of PRRSV binding exist and that entry is by endocytosis. Virus uptake in vesicles is a clathrin-dependent process, as it was blocked by the addition of cytochalasin D and co-localization of PRRSV and clathrin was found. Furthermore, by the use of two weak bases, NH4Cl and chloroquine, it was demonstrated that PRRSV uses a low pH-dependent entry pathway. In the presence of these reagents, input virions accumulated in large vacuoles, indicating that uncoating was prevented. These results indicate that PRRSV entry into AMphi involves attachment to a specific virus receptor(s) followed by a process of endocytosis, by which virions are taken into the cell within vesicles by a clathrin-dependent pathway. A subsequent drop in pH is required for proper virus replication.  (+info)

BACKGROUND: Clathrin is a multimeric protein involved in vesicle coat assembly. Recently clathrin distribution was reported to change during the cell cycle and was found to associate with the mitotic spindle. Here we test whether the recruitment of clathrin to the spindle is indicative of a critical functional contribution to mitosis. METHODOLOGY/PRINCIPAL FINDINGS: Previously a chicken pre-B lymphoma cell line (DKO-R) was developed in which the endogenous clathrin heavy chain alleles were replaced with the human clathrin heavy chain under the control of a tetracycline-regulatable promoter. Receptor-mediated and fluid-phase endocytosis were significantly inhibited in this line following clathrin knockout, and we used this to explore the significance of clathrin heavy chain expression for cell cycle progression. We confirmed using confocal microscopy that clathrin colocalised with tubulin at mitotic spindles. Using a propidium iodide flow cytometric assay we found no statistical difference in the cell
The heterotetrameric adaptor protein complex AP2 is one of the best-studied components of the endocytic machinery. The AP2 complex consists of four different subunits, α, β2, σ2, and μ2, which assemble into a core domain with two appendages (Fig. 2; Collins et al., 2002; Jackson et al., 2010). AP2 has multiple binding partners, including phosphatidylinositol 4,5-bisphosphate (PIP2), clathrin, several endocytic accessory proteins, and two signaling motifs present on some cargo receptors (see Traub, 2009 for a detailed review). The AP2 complex has classically been considered to be the master initiator of clathrin-mediated endocytosis through its role in recruiting clathrin molecules to the membrane. However, several lines of evidence question this idea.. If the AP2 complex has an essential role in initiation then its presence would be required for the formation of endocytic sites. However, in yeast the endocytosis of mating pheromone α-factor is unaffected in strains lacking functional AP2 ...
Coat proteins appear to play a general role in intracellular protein trafficking by coordinating a membrane budding event with cargo selection. Here we show that the AP-2 adaptor, a clathrin-associated coat-protein complex that nucleates clathrin-coated vesicle formation at the cell surface, can also initiate the assembly of normal polyhedral clathrin coats on dense lysosomes under physiological conditions in vitro. Clathrin coat formation on lysosomes is temperature dependent, displays an absolute requirement for ATP, and occurs in both semi-intact cells and on purified lysosomes, suggesting that clathrin-coated vesicles might regulate retrograde membrane traffic out of the lysosomal compartment. ...
To assess the role of clathrin in the bulk endocytic flow of rat foetal fibroblasts, the rate of internalization of fluid-phase and membrane-lipid tracers were compared, under control conditions and after inhibition of endocytic clathrin-coated pit formation. After intracellular potassium depletion or upon cell transfer into 0.35 M NaCl, the rate of internalization of receptor-bound transferrin and the residual membrane area of plasmalemmal clathrin-coated pits and vesicles were similarly decreased by approximately 90%. In contrast, the initial rate (, 5 min) of intracellular accumulation of the fluid-phase tracer HRP was not affected. Both in control and treated cells, the rate of HRP accumulation declined after approximately 5 min, and was twofold lower in treated cells, due to enhanced regurgitation. After correction for regurgitation, the endocytic rate constant was similar to measurements at shorter intervals and identical in control and treated cells. Similarly, the rate of internalization ...
article{1864472, abstract = {Endocytosis is a crucial mechanism by which eukaryotic cells internalize extracellular and plasma membrane material, and it is required for a multitude of cellular and developmental processes in unicellular and multicellular organisms. In animals and yeast, the best characterized pathway for endocytosis depends on the function of the vesicle coat protein clathrin. Clathrin-mediated endocytosis has recently been demonstrated also in plant cells, but its physiological and developmental roles remain unclear. Here, we assessed the roles of the clathrin-mediated mechanism of endocytosis in plants by genetic means. We interfered with clathrin heavy chain (CHC) function through mutants and dominant-negative approaches in Arabidopsis thaliana and established tools to manipulate clathrin function in a cell type-specific manner. The chc2 single mutants and dominant-negative CHC1 (HUB) transgenic lines were defective in bulk endocytosis as well as in internalization of ...
EpsinR is a clathrin-coated vesicle (CCV) enriched 70-kD protein that binds to phosphatidylinositol-4-phosphate, clathrin, and the gamma appendage domain of the adaptor protein complex 1 (AP1). In cells, its distribution overlaps with the perinuclear pool of clathrin and AP1 adaptors. Overexpression disrupts the CCV-dependent trafficking of cathepsin D from the trans-Golgi network to lysosomes and the incorporation of mannose-6-phosphate receptors into CCVs. These biochemical and cell biological data point to a role for epsinR in AP1/clathrin budding events in the cell, just as epsin1 is involved in the budding of AP2 CCVs. Furthermore, we show that two gamma appendage domains can simultaneously bind to epsinR with affinities of 0.7 and 45 microM, respectively. Thus, potentially, two AP1 complexes can bind to one epsinR. This high affinity binding allowed us to identify a consensus binding motif of the form DFxDF, which we also find in gamma-synergin and use to predict that an uncharacterized EF-hand
The early events of viral infection usually involve the attachment of virus to cellular receptor molecules on the plasma membrane of host cells. This is followed by internalization, uncoating, and subsequent virus gene transcription and/or translation at specific locations in cells. Studies have clearly demonstrated that animal viruses can utilize different internalization and trafficking pathways that allow specific localization within the cells upon entry for a successful infection (15). For enveloped viruses, the entry process can occur either via the fusion of virus envelope glycoproteins at the plasma membrane at neutral pH to promote the internalization of viral nucleocapsids or virus particles undergoing endocytosis prior to fusion with endocytic membrane. For the latter, conformational change of the virus fusion protein to expose the hydrophobic fusion peptide is induced by an acidic pH for the release of the viral nucleocapsids into the cytoplasm (14).. In the present study, a variety ...
Link to Pubmed [PMID] - 20486136. Bioessays 2010 Jun;32(6):496-504. Clathrin and the endocytosis machinery has recently been described as being required in mammalian cells for the internalization of large particles including pathogenic bacteria, fungi, and large viruses. These apparently unexpected observations, within the framework of the classical mechanisms for the formation of clathrin-coated vesicles, are now considered as examples of a new non-classical function of clathrin, which can promote the internalization of membrane domains associated to planar clathrin lattices. The role of actin downstream of clathrin seems to be critical for this still poorly characterized process. The historical frontier between endocytosis and phagocytosis is vanishing in the light of this new role for clathrin.. http://www.ncbi.nlm.nih.gov/pubmed/20486136 ...
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Our study provides evidence for a new role of Src in the phosphorylation of the clathrin adaptor complex AP-2 (via the phosphorylation of the β-subunit, i.e. β2-adaptin). We identify Y737 in the ear domain of β2-adaptin as an important Src target. We show that this residue represents a regulatory site for controlling the dissociation of β-arrestin from AP-2 in clathrin-coated pits (CCPs). Our results not only extend the pleiotropic function of Src in GPCR internalization, but also highlight the importance of receptor-dependent signaling in the clathrin-mediated endocytosis of AT1R.. β-arrestins are multifunctional adaptors involved in the endocytosis and signaling of many GPCRs (Claing et al., 2002; Lefkowitz, 1998). They have been shown to target receptors to CCPs through binding to clathrin and the β-subunit of AP-2 (Goodman, Jr et al., 1996; Laporte et al., 1999), and to act as signal transducers by recruiting different kinases to GPCRs (Luttrell and Luttrell, 2004). We have previously ...
VOLUME 21 ISSUE 12 12 2011 1655 1661 Novel functions of endocytic player clathrin in mitosis Wenxiang Fu Qing Jiang and Chuanmao Zhang The MOE Key Laboratory of Cell Proliferation and Differentiation and the State Key Laboratory of Bio membrane and Membrane Biotechnology College of Life Sciences Peking University Beijing 100871 China Correspondence Chuanmao Zhang Tel 86 10 62757173 E mail zhangcm pku edu cn Clathrin has been widely recognized as a pivotal player in endocytosis in which several adaptors and accessory proteins are involved Recent studies suggested that clathrin is also essential for cell division Here this review mainly focuses on the clathrin dependent mechanisms involved in spindle assembly and chromosome alignment In mitosis clathrin forms a complex with phosphorylated TACC3 to ensure spindle stability and proper chromosome alignment The clathrin regulated mechanism in mitosis requires the crosstalk among clathrin spindle assembly factors SAFs Ran GTP and mitotic kinases ...
The structure of Clathrin is known as a triskeleion structure with there being three bent legs extending from a central point known as the central trimerization domain. Clathrin is made up of six chains of protein that are braided together in such a way to form to form its distinct shape. Three of these chains are known as heavy chains and form the backbone of Clathrin, they consist of two sub domains a N-terminal zone and a proximal leg domain. A seven bladed β-propeller structure is what the N-terminal domain consists off. While the proximal leg consists of a super Helix (Conner and Schmid, 2002). The other three chains are known as light chains and regulate formation and disassembly of the Clathrin and can be found connected to the proximal portion of the heavy chains. Multiple Clathrin molecules have the ability to form a variety of complex shapes when they interact, they can form 5 or 6 sided rings with the 5 sided kind having a greature curvature and when enough get together they can form ...
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The lipid mediator sphingosine 1-phosphate (S1P) regulates a wide range of cellular activities, including vascular maturation, angiogenesis, and immune-cell trafficking. Among the five known receptors for S1P (S1PR1-S1PR5), S1PR1 is a critical regulator of lymphocyte trafficking: its signaling is required for lymphocyte egress from lymphoid organs, while its down-modulation by agonist-induced internalization is a prerequisite for lymphocyte entry into lymphoid organs from the bloodstream. Despite the importance of S1PR1 down-regulation in determining lymphocyte behavior, the molecular mechanism of its internalization in lymphocytes has not been defined. Here we show that agonist-induced S1PR1 internalization in T cells occurs via clathrin-mediated endocytosis and is regulated by moesin, an ezrin-radixin-moesin (ERM) family member. In S1P-stimulated T cells, S1PR1 relocalized within clathrin-coated vesicles (CCVs) and early endosomes, and S1PR1 internalization was blocked when clathrin was
The lectin isolated from Xerocomus chrysenteron (XCL) displays a toxic activity towards insects. In order to assess its possible mode of action and to gather useful data for its potential use in insect-resistant transgenic plants, we investigated the
We found that lipid vesicles are internalized by both clathrin-mediated and clathrin-independent endocytosis pathways which require an acidification step for liposomes destabilization and fusion with the endosomes. The fusion event possibly triggers a microtubule driven pathway which avoids classical sorting endosomes and favours an ER pathway. This is an alternate to the SV40 virus caveolar route using a pH dependent, direct plasma membrane-to-ER pathway to efficiently deliver extracellular encapsulated cargo to the ER. Whilst further studies are needed to dissect this novel pathway in more detail, the finding of a second major ER targeting route leads to the idea that the import of molecules from the outside cell into the ER could be as important for the cell function as its secretion processes ...
Clathrin-mediated endocytosis (CME) is a key metabolic pathway that plays a central role in the delivery of nutrients and drug carriers into cells. In this study, we model the interactions of lipid membranes with different types of protein scaffolds and active forces to provide mechanistic insights into CME. To this end, we develop and employ an extended theoretical framework of lipid membranes that entertains spatial heterogeneity and local anisotropy that could arise from membrane-protein interactions. We show that a departure from homogeneity and isotropy can lead to a variable surface tension field, conventionally assumed to be a constant parameter. We model the impact of resting tension in a cell and discuss its consequences on the minimal protein machinery needed to complete vesicle formation. Based on our quantitative model and findings, we highlight the physical principles that unify CME in apparently distinct yeast and mammalian cells.
SMAP2 immunoprecipitated clathrin and AP-1 through a putative clathrin-binding domain and a CALM-binding domain, and SMAP2 mutants that did not interact with clathrin or AP-1 could not localize to recycling ...
mmmmm cow brains the concept centers on a particular protein called clathrin which has a unique knack for assembling itself into versatile structures that foster the formation of complex molecules clathrin is present in every cell in the human body but cows possess a vast wealth of it in their bovine brains that make them an ideal source for the stuff and given the right biochemical directions researchers think they can coax clathrin into creating better batteries and solar cells http www popsci com science article - protein-cow-brains-could-build-better-batteries-solar-cells
Rabbit Polyclonal Anti-Clathrin interactor 1 Antibody. Validated: WB, ICC/IF, IHC, IHC-P. Tested Reactivity: Human, Mouse, Rat. 100% Guaranteed.
Reactome is pathway database which provides intuitive bioinformatics tools for the visualisation, interpretation and analysis of pathway knowledge.
Control of E-cadherin apical localisation and morphogenesis by a SOAP-1/AP-1/clathrin pathway in C. elegans epidermal cells. ...
Thermodynamics of protein-mediated membrane deformation â application to clathrin dependent and clathrin independent endocytosis (2009 ...
Reactome is pathway database which provides intuitive bioinformatics tools for the visualisation, interpretation and analysis of pathway knowledge.
SSO mediated functional uptake in MHT cells is AP2M1 dependent and clathrin independent. (A) MHT cells were treated with 25 nM control, AP2M1, clathrin or RNA
Function: Phosphorylates the AP2M1/mu2 subunit of the adaptor protein complex 2 (AP-2). May play a role in regulating aspects of clathrin-mediated endocytosis (By similarity ...
The vesicle simulation is available in two forms, one having enough for about one and a half vesicles (vesicle.obj) and the other having enough for about three (vesicle2.obj). Both can use either the vesicle_hub.tmpl or the vesicle_no_hub.tmpl template files (the first contains a sphere at the centre of each clathrin and is more polished visually, the second is faster). Note that in some runs of the simulation many fragments of vesicles form, rather than the few complete structures ...
Mills IG, Praefcke GJ, Vallis Y, Peter BJ, Olesen LE, Gallop JL, Butler PJ, Evans PR, McMahon HT. EpsinR: an AP1/clathrin interacting protein involved in vesicle trafficking. J Cell Biol. 2003 Jan 20;160(2):213-22. Epub 2003 Jan 21. PMID:12538641 doi:http://dx.doi.org/10.1083/jcb.200208023 ...
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Cell-surface viral proteins most frequently enter the cell through clathrin or caveolae endocytosis. Respiratory syncytial virus antigen internalization by immune cells is via caveolin, however, uptake of paramyxovirus cell membrane proteins by non-immune cells is done through clathrin-coated pits. In this work, the uptake of respiratory syncytial virus cell surface glycoproteins by non-immune human epithelial cells was investigated through indirect immunofluorescence with polyclonal anti-RSV antibody and confocal lasser-scanner microscopy. Clathrin and caveolae internalization pathways were monitored through specific inhibitors monodansylcadaverine (MDC) and methyl-beta-cyclodextrin (MBCD), respectively. Internalization of RSV antigens was inhibited by MDC but not by MBCD, implying that clathrin-mediated endocytosis is the major uptake route of RSV antigens by an epithelial human cell line.
In this report we have addressed the biological consequence of endofin-mediated TOM1 recruitment onto endosomes. Our results collectively suggest that TOM1 serves as an adaptor for endofin to recruit clathrin heavy chain onto the endosomes. This conclusion is supported by several lines of evidence as described in our study.. First, via large-scale pull-down experiments using immobilized GST-TOM1(300-492), we have recovered clathrin heavy chain as the major and specific partner for TOM1. This conclusion was corroborated by analytical pull-down experiments showing that clathrin heavy chain was very efficiently retained by immobilized GST-TOM1(300-492), so much so that it was depleted from the cytosol. The specific interaction between TOM1 and clathrin heavy chain was further defined by our identification of three sites in the carboxyl-terminal region of TOM1, which seem to act together for efficient interaction with clathrin. Moreover, the specific blockage of interaction between TOM1 and clathrin ...
Clathrin, a three-legged triskelion composed of three clathrin heavy chains (CHCs) and three light chains (CLCs), plays a critical role in clathrin-mediated endocytosis (CME) in eukaryotic cells. In this study, the genes ZmCHC1 and ZmCHC2 encoding clathrin heavy chain in maize were cloned and characterized for the first time in monocots. ZmCHC1 encodes a 1693-amino acid-protein including 29 exons and 28 introns, and ZmCHC2 encodes a 1746-amino acid-protein including 28 exons and 27 introns. The high similarities of gene structure, protein sequences and 3D models among ZmCHC1, and Arabidopsis AtCHC1 and AtCHC2 suggest their similar functions in CME. ZmCHC1 gene is predominantly expressed in maize roots instead of ubiquitous expression of ZmCHC2. Consistent with a typical predicted salicylic acid (SA)-responsive element and four predicted ABA-responsive elements (ABREs) in the promoter sequence of ZmCHC1, the expression of ZmCHC1 instead of ZmCHC2 in maize roots is significantly up-regulated by SA or ABA,
Assembly protein recruiting clathrin and adapter protein complex 2 (AP2) to cell membranes at sites of coated-pit formation and clathrin-vesicle assembly. May be required to determine the amount of membrane to be recycled, possibly by regulating the size of the clathrin cage. Involved in AP2-dependent clathrin-mediated endocytosis at the neuromuscular junction. Plays a crucial role in fetal and adult hematopoiesis, and normal prenatal and postnatal growth and viability.
Fingerprint Dive into the research topics of CAP-1A is a novel linker that binds clathrin and the voltage-gated sodium channel Na,sub,v,/sub,1.8. Together they form a unique fingerprint. ...
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Clathrin-coated vesicles are the most prominent carriers of membrane traffic from cell surface to endosomes (endocytosis), a pathway by which hormones, transferrin, immunoglobulins, LDL, viruses, and their receptors enter cells. They are also important for traffic between endosomes and the trans-Golgi network. In this presentation, I will discuss (i) technological and analytical advances that I developed to directly visualize clathrin-mediated membrane traffic in three dimensions and in living cells; (ii) data obtained using these advances that defined a role for actin filament polymerization in counteracting membrane tension during clathrin-coated vesicle budding at the apical surface of polarized epithelial cells; and (iii) how these advances can be used to study a wide variety of biological processes that occur in living cells and tissues. ...
During clathrin-mediated endocytosis, it has been thought that the sensing and binding of the clathrin adaptor protein AP2 to cargo and lipids leads to the recruitment of clathrin, nucleating the formation of a clathrin-coated pit. Henne et al. have now found that this process of AP2 binding may not in fact represent either the first or the nucleation event of endocytosis. Instead, ubiquitous proteins called FCHo1/2 (F-BAR proteins) bind to the plasma membrane and define the sites of endocytosis independently of AP2. The F-BAR protein can generate very low curvatures and, at higher concentrations, generates higher curvatures like those required at the neck of budding vesicles. The C terminus of the protein has a μ-homology domain (with homology to the μ domain of the AP2 complex) that interacts with Eps15 and intersectin and via these proteins recruits AP2, which further recruits clathrin. Thus, a curvature-inducing protein can act to nucleate clathrin-coated pit assembly during ...
Steps in CCV assembly and links to structures and information around clathrin-coated vesicle formation and other forms of vesicle budding
PICALM, the gene encoding phosphatidylinositol-binding clathrin assembly (picalm) protein, was recently shown to be associated with risk of Alzheimer disease (AD). Picalm is a key component of clathrin-mediated endocytosis. It recruits clathrin and adaptor protein 2 (AP-2) to the plasma membrane and, along with, AP-2 recognizes target proteins. The attached clathrin triskelions cause membrane deformation around the target proteins enclosing them within clathrin-coated vesicles to be processed in lysosomes or endosomes. We examined the distribution of picalm in control and AD brain tissue and measured levels of picalm messenger RNA (mRNA) by real-time polymerase chain reaction. Immunolabeling of brain tissue showed that picalm is predominately present in endothelial cells. This was further supported by the demonstration of picalm in human cerebral microvascular cells grown in culture. Picalm mRNA was elevated in relation to glyceraldehyde-3-phosphate dehydrogenase but not factor VIII-related ...
Toxoplasma gondii possesses a highly polarized secretory system, which efficiently assembles de novo micronemes and rhoptries during parasite replication. These apical secretory organelles release their contents into host cells promoting parasite invasion and survival. Using a CreLox-based inducible knock-out strategy and the ddFKBP over-expression system, we unraveled novel functions of the clathrin adaptor complex TgAP1. First, our data indicate that AP1 in T. gondii likely functions as a conserved heterotetrameric complex composed of the four subunits γ, β, μ1, σ1 and interacts with known regulators of clathrin-mediated vesicular budding such as the unique ENTH-domain containing protein, which we named Epsin-like protein (TgEpsL). Disruption of the μ1 subunit resulted in the mis-sorting of microneme proteins at the level of the Trans-Golgi-Network (TGN). Furthermore, we demonstrated that TgAP1 regulates rhoptry biogenesis by activating rhoptry protein exit from the TGN, but also ...
Clathrin-mediated endocytosis is a critical process through which a wide variety of extracellular material is internalized. The primary component, clathrin, forms a cargo-selective lattice at the plasma membrane, as well as on endosomes and the TGN, though the cargo-selective components are incompletely defined. An ideal tool for understanding the spatio-temporal dynamics of both the clathrin coat and the cargo selected is total internal reflection fluorescence microscopy (TIR-FM), which permits selective imaging of events closely apposed to the ventral plasma membrane. Previously, observation of the clathrin coat has shown both static and dynamic populations, with some dynamic structures undergoing microtubule-dependent motion; the 70-110 nm decay constant of the TIR-FM field has led to the assumption that these are all representative of coated pits. Here, I demonstrate that the dynamic population of clathrin is primarily endosomal, as it lacks colocalization with the plasma membrane-specific ...
Bicaudal-D (Bic-D), Egalitarian (Egl), microtubules and their motors form a transport machinery that localizes a remarkable diversity of mRNAs to specific cellular regions during oogenesis and embryogenesis. Bic-D family proteins also promote dynein-dependent transport of Golgi vesicles, lipid droplets, synaptic vesicles and nuclei. However, the transport of these different cargoes is still poorly understood. We searched for novel proteins that either mediate Bic-D-dependent transport processes or are transported by them. Clathrin heavy chain (Chc) co-immunopurifies with Bic-D in embryos and ovaries, and a fraction of Chc colocalizes with Bic-D. Both proteins control posterior patterning of the Drosophila oocyte and endocytosis. Although the role of Chc in endocytosis is well established, our results show that Bic-D is also needed for the elevated endocytic activity at the posterior of the oocyte. Apart from affecting endocytosis indirectly by its role in osk mRNA localization, Bic-D is also ...
PhD Project - High resolution cryo-electron microscopy of clathrin cage networks at University of Warwick, listed on FindAPhD.com
Endocytosis is an essential phenomenon in eukaryotic cells. In animal cells, dynamin and clathrin play central roles in vesicle formation in the process of endocytosis, but the roles of similar proteins in plants are less well understood. Here, we observed the localization pattern and behavior of GFP-labeled ,i,Arabidopsis,/i, dynamin-related proteins (DRP1A and DRP2B), and clathrin light chain (AtCLC) around the plasma membrane in tobacco suspension cells by using variable incidence angle fluorescence microscopy (VIAFM). GFP fusions of DRP1A, DRP2B and AtCLC were observed as dot-like puncta 200-500 nm in diameter. The puncta moved to and away from the cell surface or also assembled and disassembled. The localization pattern and behavior of the puncta were similar to those of animal dynamin and clathrin signals reported previously. These results raise the possibility that DRP1A, DRP2B and AtCLC are involved in membrane trafficking around the plasma membrane, including endocytosis.. ...
The general objective of our lab is to understand the functions of clathrin-coated structures (CCSs) during the different steps of cancer development. CCSs recruit specific cell surface receptors and progressively shape the plasma membrane in receptor-containing vesicles that are released in the cytosol. This endocytosis machinery allows for nutrient uptake but also for the fine-tuned control of signaling pathways triggered by cell surface receptors. As a consequence, deregulation of endocytosis has been linked to many pathological situations, including cancers.. Tumor development is accompanied by dramatic changes in the mechanical characteristics of tissues. Also, when cancer cells invade the stroma to establish distant metastases, they migrate in an environment with different topological features than the tumor mass. However, it is not known how the physical parameters of the environment impact on CCSs and what are the consequences for the cell.. Our team addresses this general question by ...
Immunogen = synthetic peptide: E E D P A A A F L A Q Q E S E I A G I E N D, corresp. to amino acids 23-44 of Cow Clathrin light chain. ...
Zwiewka, M; Nodzynski, T; Robert, S; Vanneste, S; Friml, J, 2015: Osmotic Stress Modulates the Balance between Exocytosis and Clathrin-Mediated Endocytosis in Arabidopsis thaliana. MOLECULAR PLANT 8(8), p. 1175 - 1187, doi: 10.1016/j.molp.2015.03.007. Research Groups:. ...
Glyvuk et al (2010) argue that the CME of SV membranes represents a kinetic bottleneck of the recycling pathway. Under conditions of sustained activity BE provides a compensatory mechanism to balance high exocytic load with matching endocytic activity. Vacuolar membrane invaginations are then consumed by undefined budding events that chop these membranes into small vesicles that may re‐enter the SV cycle (Figure 1A). It is this consumption step that the authors envision to depend on AP‐1/σ1B (Figure 1B). The experimental evidence for this model at present remains indirect. AP‐1, as its relative AP‐2, is one of the major recruitment factors for clathrin and loss of either protein complex results in depletion of clathrin‐coated pits from TGN/endosomes or the plasmalemma, respectively. Why then do AP‐1/σ1B‐KO mice accumulate clathrin‐coated pits on endosome‐like vacuoles? One possibility is that other σ1 isoproteins such as σ1A do a poor job in functionally replacing σ1B on ...
Researchers of the group of cellular and molecular neurobiology of the Bellvitge Biomedical Research Institute and the University of Barcelona, led by researcher Artur Llobet, have shown that synaptic levels of the protein clathrin are a determinant factor for synaptic plasticity of neurons.
Clathrin (see MIM 118955)-mediated endocytosis is a major mechanism for internalization of proteins and lipids. Members of the connecdenn family, such as DENND1A, function as guanine nucleotide exchange factors (GEFs) for the early endosomal small GTPase RAB35 (MIM 604199) and bind to clathrin and clathrin adaptor protein-2 (AP2; see MIM 601024). Thus, connecdenns link RAB35 activation with the clathrin machinery (Marat and McPherson, 2010 [PubMed 20154091]).[supplied by OMIM, Nov 2010 ...
Video articles in JoVE about clathrin coated vesicles include Applications of pHluorin for Quantitative, Kinetic and High-throughput Analysis of Endocytosis in Budding Yeast, Visualizing Clathrin-mediated Endocytosis of G Protein-coupled Receptors at Single-event Resolution via TIRF Microscopy, Measuring Synaptic Vesicle Endocytosis in Cultured Hippocampal Neurons, The Cell-based L-Glutathione Protection Assays to Study Endocytosis and Recycling of Plasma Membrane Proteins, Pulling Membrane Nanotubes from Giant Unilamellar Vesicles, In vivo and in vitro Studies of Adaptor-clathrin Interaction, Models and Methods to Evaluate Transport of Drug Delivery Systems Across Cellular Barriers, Methods for Cell-attached Capacitance Measurements in Mouse Adrenal Chromaffin Cell, Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes, Nitrogen Cavitation and Differential Centrifugation Allows for Monitoring the
Component of the adaptor complexes which link clathrin to receptors in coated vesicles. Clathrin-associated protein complexes are believed to interact with the cytoplasmic tails of membrane proteins, leading to their selection and concentration. AP50 is a subunit of the plasma membrane adaptor. The complex binds polyphosphoinositide-containing lipids. [-] ...
Expression of α-Synuclein and clathrin in Z310 cells with or without a-Syn treatment in a typical experiment (n = 5). Z310 cells are immortalized rat choro
Ybe and Niu used X-ray crystallography to look at an area of interest on the surface of HIP1, which works in concert with clathrin to traffic nutrients into a cell, and has long been implicated as playing an important role in the development of Huntingtons disease. They learned that the potential binding surface of HIPPI in HIP1 has an unexpected shape for a binding site, a spiraling spiral called a "coiled coil." This finding was contrary to predicted results that the binding surface that receives HIPPI is folded into a so-called death effector domain. Using the information from the published molecular structure of HIP1, IU biologists hope to be able to test which protein connections are ultimately responsible for triggering the chain of interactions leading to Huntingtons disease and how to block them. For example, they observed that clathrin, protein involved in bringing nutrients to the cell, binds with HIP1 right next door to where HIPPI binds. While clathrin "packages" nutrients for a ...
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Coated pit: …of the membrane called a coated pit, which is lined by a special protein known as clathrin. As the coated pit invaginates, it is pinched off in the cytoplasm to form a coated vesicle. The coated vesicle fuses with cytoplasmic endosomes (membrane-enclosed vesicles) and then with cell organelles called lysosomes,…
Abstract In metazoans, intracellular trafficking is mediated by many conserved pathways. The main trafficking pathways involve the Adaptor Protein complexes (AP) AP-1, AP-2 and AP-3, Clathrin and Clathrin Associated Sorting Proteins (CLASPS). These proteins direct cargoes to and from the Golgi, the endosome, the lysosome and the plasma membrane. Lipoprotein receptors are one class of proteins that use these pathways and are endocytosed by AP-2 and Disabled family proteins. Disabled family proteins are ~ conserved family of monomeric adaptor proteins and consist of a single PTB domain and have a variety of motifs associated with trafficking. C. elegans has one Disabled family member, DAB-1.1 show here that .DAB-1 is a generally expressed regulator of membrane trafficking. Loss of dab-1 function, as well as an array of molecules involved membrane trafficking, perturbs cadherin-catenin function, though I have not been able to determine the precise basis of these genetic interactions. I show that ...
Involved in cell growth regulation. May be involved in the regulation of mitogenic signals and control of cell proliferation. Involved in the internalization of ligand-inducible receptors of the receptor tyrosine kinase (RTK) type, in particular EGFR. Plays a role in the assembly of clathrin-coated pits (CCPs). Acts as a clathrin adapter required for post-Golgi trafficking. Seems to be involved in CCPs maturation including invagination or budding. Involved in endocytosis of integrin beta-1 (ITGB1) and transferrin receptor (TFR); internalization of ITGB1 as DAB2-dependent cargo but not TFR seems to require association with DAB2. ...
In light of the presented data, we suggest that LRRK2 is part of a functional protein network that controls SV trafficking within the recycling pool by interacting with a subset of presynaptic proteins. A role of LRRK2 in vesicle trafficking involving Rab5b had already been suggested (Shin et al., 2008), but we can now show for the first time that electrophysiological properties as well as vesicular trafficking in the presynaptic pool depend on the presence of LRRK2 as an integral part of presynaptic protein complex. We have in fact identified presynaptic proteins-NSF, AP-2 complex subunits, SV2A, synapsing, and syntaxin 1-as well as actin as putative LRRK2 interactors. These proteins have been described previously as key elements of synaptic vesicle trafficking (Takamori et al., 2006). NSF catalyzes the release of the SNARE complex (SNAP-25, syntaxin 1, and Vamp) and allows the first step of the endocytic cycle (Littleton et al., 1998, 2001). The clathrin complex [clathrin, AP-2 adaptor ...
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The interaction between cytoplasmic coat proteins and specific signals in the cytoplasmic domains of integral membrane proteins is a general mechanism controlling protein sorting. The best-characterized sorting signal is the tyrosine-based sorting motif that is involved in various sorting events by interaction with adaptor proteins (Trowbridge et al., 1993; Mellman, 1996; Marks et al., 1997). The YRSLE sequence present in the cytoplasmic domain of the neuronal form of L1 conforms to the tyrosine-based sorting motif. We have found that the AP-2 adaptor specifically recognizes and interacts with the YRSLE sequence, resulting in clathrin-mediated endocytosis of L1 (H. Kamiguchi, K. E. Long, M. Pendergast, A. W. Schaefer, I. Rapoport, T. Kirchhausen, and V. Lemmon, unpublished observations). These observations indicate that the YRSLE sequence of L1 actually functions as a tyrosine-based sorting signal. The L1 cytoplasmic domain contains three other tyrosine residues (Hlavin and Lemmon, 1991), ...
In the second lecture, the next steps in viral infection are described. Endocytosis of plasma membrane bound viruses can occur via a number of mechanisms including caveolar, clathrin, non-clathrin, or lipid raft mediated pathways. The internalized vi
Plasmid pSNAP-CLC-SNAP from Dr. Xiaowei Zhuangs lab contains the insert Clathrin, light polypeptide (LCa) and is published in Nat Methods. 2011 Jun;8(6):499-508. Epub 2011 May 8. This plasmid is available through Addgene.
Plasmid mEos4a-Clathrin-15 from Dr. Michael Davidsons lab contains the insert Clathrin. This plasmid is available through Addgene.
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Fur balls occur when cats clean themselves, ingesting their own fur. While these are common, be sure to brush your cats coat on a regular basis so that this doesnt lead to serious, and even fatal, problems. read more ...
The animation shows calcium-stimulated exocytosis of synaptic vesicles followed by clathrin-mediated vesicle recycling. Many of the molecular components that are involved in synaptic vesicle priming, docking, fusion, and endocytosis are shown. Although the process in the animation describes synaptic vesicle cycling, similar cellular processes occur for most calcium-coupled secretory and clathrin-mediated endocytotic events. The animation could be used to help illustrate the sequence of events associated with both exocytosis and endocytosis, as well as aid in understanding the processes involved in neurotransmitter release in response to nerve stimulation.. [Resource Details] ...
The animation shows calcium-stimulated exocytosis of synaptic vesicles followed by clathrin-mediated vesicle recycling. Many of the molecular components that are involved in synaptic vesicle priming, docking, fusion, and endocytosis are shown. Although the process in the animation describes synaptic vesicle cycling, similar cellular processes occur for most calcium-coupled secretory and clathrin-mediated endocytotic events. The animation could be used to help illustrate the sequence of events as. Published by Learning Registry #GoOpen. 4 Views, 0 Likes on Docs.com. #synapse #neuron #NSDL #NSDL_SetSpec_BEN #movie #signal transduction #endocytosis #Life Science
Adapter protein that functions as clathrin-associated sorting protein (CLASP) required for clathrin-mediated endocytosis of selected cargo proteins. Can bind and assemble clathrin, and binds simultaneously to phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) and cargos containing non-phosphorylated NPXY internalization motifs, such as the LDL receptor, to recruit them to clathrin-coated pits. Can function in clathrin-mediated endocytosis independently of the AP-2 complex. Involved in endocytosis of integrin beta-1; this function seems to redundant with the AP-2 complex and seems to require DAB2 binding to endocytosis accessory EH domain-containing proteins such as EPS15, EPS15L1 and ITSN1. Involved in endocytosis of cystic fibrosis transmembrane conductance regulator/CFTR. Involved in endocytosis of megalin/LRP2 lipoprotein receptor during embryonal development. Required for recycling of the TGF-beta receptor. Involved in CFTR trafficking to the late endosome. Involved in several receptor-mediated
Metastasis is a multistep process requiring cancer cell signaling, invasion, migration, survival, and proliferation. These processes require dynamic modulation of cell surface proteins by endocytosis. Given this functional connection, it has been suggested that endocytosis is dysregulated in cancer. To test this, we developed In-Cell ELISA assays to measure three different endocytic pathways: clathrin-mediated endocytosis, caveolae-mediated endocytosis, and clathrin-independent endocytosis and compared these activities using two different syngeneic models for normal and oncogene-transformed human lung epithelial cells. We found that all endocytic activities were reduced in the transformed versus normal counterparts. However, when we screened 29 independently isolated non-small cell lung cancer (NSCLC) cell lines to determine whether these changes were systematic, we observed significant heterogeneity. Nonetheless, using hierarchical clustering based on their combined endocytic properties, we ...
The present invention relates in a first aspect to a method of coating surfaces of substrates with a lattice-like structure. In particular, the present invention relates to an in vitro method of coating surfaces by binding of epsin or a fragment thereof on the surface and, thereafter, binding of a compound forming the lattice like structure, in particular, binding of the clathrin heavy chain, to the epsin bound on the surface, thus, obtaining a coated substrate having a lattice like structure on the surface. In another aspect, the present invention relates to an in vitro method of producing nanometer-sized liposomes having a clathrin structure on its surface. In addition, substrates, like elements or devices, with coated surfaces having a lattice-like structure on the surface are provided obtainable by a method according to the present invention.
UNC-51 overexpression can inhibit the transferrin endocytosis in the transfected COS-7 cells. We think that this inhibition of transferrin endocytosis should be caused by UNC-51-dependent...
Background: Epsins are a family of ubiquitin-binding endocytic clathrin adaptors. We recently published that endothelial epsins function as critical regulators of tumor angiogenesis by controlling VEGF signaling (JCI, 2012; ATVB, 2013). Our goal is to define the novel role of epsins in macrophages in regulating atherogenesis.. Methods and Results: We engineered mice with specific deletion of epsins in myeloid cells (MΦ-DKO). Strikingly, MΦ-DKO mice on ApoE-/- background fed western diet significantly reduced atherosclerotic lesion formation and foam cell accumulation. In macrophages, epsin deficiency did not alter LDL scavenger receptors, CD36, Lox1 or SRB1, or reverse cholesterol transport proteins, ABCA1 or ABCG1, but did significantly reduce Lucifer Yellow pinocytosis, indicating a major defect in lipid uptake. Epsin deficiency did decrease total and surface protein levels of LRP-1, a protein with anti-inflammatory and anti-atherosclerotic properties. Oil Red O staining of isolated ...
Nature, 417(6888): 555-559. Vesicle pools Vesicle mobilization, docking, priming and fusion Three endocytic pathways The dynamic time course of endocytosis The time course of endoc3l;osis is regulated by stimulation Mechanisms that may regulate the time course of endocytosis Clathrin-dependent versus clathrin-independent endocytosis Regulation of endocytosis by single rate-limiting factors The effects of calcium on endocytosis Hypotheses that may account for calcium-mediated facilitation of endocytosis The functional significance of regulation of endoc3l;osis The contribution of slower endocytosis to short-term synaptic depression The maintenance of transmitter release by retrieving vesicles into the reserve pool and/or the readily releasable pool Is endocytosis fast enough to limit transmitter release from the fusion pore? Waldeck RF, Pereda A, Faber DS. 2000. Properties and plasticity of paired-pulse depression at a central synapse. J Neurosci, 20: 5312-5320. Wang LY, Kaczmarek LK. 1998. ...
DUGi: Viewing Item from repository DUGiDocs: Peptide conjugates incorporating the red-ox active ligands Me2PyTACN or (S,S)-BPBP at the N- or the C-terminus of the cell-penetrating peptide BP16 were synthesized (PyTACN-BP16 (BP341), BP16-PyTACN (BP342), BPBP-BP16 (BP343), and BP16-BPBP (BP344)). Metal binding peptides bearing at the N-terminus the ligand, an additional Lys and a β-Ala were also prepared (PyTACN-βAK-BP16 (BP345) and BPBP-βAK-BP16 (BP346)). Moreover, taking into account the clathrin-dependent endocytic mechanism of BP16, the enzymatic cleavable tetrapeptide Gly-Phe-Leu-Gly was incorporated between the ligand and the N- or C-terminus of BP16 (BPBP-GFLG-BP16 (BP347) and BP16-GLFG-BPBP (BP348). Analysis of the cytotoxicity of all the peptide conjugates showed that: (i) the position of the ligand influenced the IC50 values, (ii) the incorporation of the βAla-Lys dipeptide rendered non active sequences, (iii) peptide conjugates derived from the (S,S)-BPBP ligand were more active than
This gene encodes a protein associated with the cytoplasmic surface of synaptic vesicles. A subset of patients with stiff person syndrome who were also affected by breast cancer are positive for autoantibodies against this protein. Alternate splicing of this gene results in two transcript variants encoding different isoforms. Additional splice variants have been described, but their full length sequences have not been determined.[6] Amphiphysin is a brain-enriched protein with an N-terminal lipid interaction, dimerisation and membrane bending BAR domain, a middle clathrin and adaptor binding domain and a C-terminal SH3 domain. In the brain, its primary function is thought to be the recruitment of dynamin to sites of clathrin-mediated endocytosis. There are 2 mammalian amphiphysins with similar overall structure. A ubiquitous splice form of amphiphysin 2 that does not contain clathrin or adaptor interactions is highly expressed in muscle tissue and is involved in the formation and stabilization ...
The protein encoded by this gene is the medium chain of the trans-Golgi network clathrin-associated protein complex AP-1. The other components of this complex are beta-prime-adaptin, gamma-adaptin, and the small chain AP1S1. This complex is located at the…
AP2-associated protein kinase 1 (Aak1) is a member of the Ark1/Prk1 subfamily of Ser/Thr protein kinases that are thought to regulate endocytosis by phosphorylating the accessory endocytic components. Aak1 interacts with and phosphorylates the mu2 subunit of the AP-2 complex, which promotes binding of the AP-2 to tyrosine based (Yxxf) internalization motif-containing receptors and subsequent receptor endocytosis. At least two isoforms of Aak1 are known to exist; the longer isoform contains an extended carboxy-terminus that contains an additional clathrin-binding domain. Overexpression of this long isoform or Aak1 depletion by RNA interference impairs transferrin recycling from the early/sorting endosome, suggesting that Aak1 functions at multiple steps of the endosomal pathway by regulating transferrin internalization and its recycling back to the plasma membrane. ...
Katya Heldwein, PhD, Principal Investigator. Katya received her PhD from Oregon Health Sciences University in Portland, OR where she studied ligand recognition by bacterial transcription regulators using x-ray crystallography in the laboratory of Richard Brennan. She then did her postdoctoral work at Harvard Medical School in the laboratory of Stephen Harrison where she initially worked on clathrin adaptor complexes and later delved into herpesvirus cell entry. She opened her own laboratory at Tufts University School of Medicine in the Fall of 2006.. ...
It is known for decades, and most recently seen in dynamin knockout mice, that endocytosis continues when classical mechanisms are blocked or deleted. With the help of ubiquitin, yeast live without clathrin and fungi reveal still other alternatives in the absence of clathrin. We discovered a few years ago that 50 percent of the cell surface of many cells can be removed in seconds by Ca-dependent endocytosis that requires no classical endocytic player. This response is unrelated to either apoptosis or autophagy; cell cultures survive and even flourish in the wake of this remodeling. This form of endocytosis, relying on no known adapter or cytoskeleton, can quantitatively remove PD-1 receptors from the T-cell surface membrane, receptors whose inactivation is a key to activating immune responses. Thus, domain-dependent endocytosis can be concentrative as well as contributing to overall fluid-phase endocytosis. As for exocytosis, Ca stress-dependent exocytosis can rapidly increase membrane area by ...
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Ford, M.G.J., Mills, I.G., Peter, B.J., Vallis, Y., Praefcke, G.J.K., Evans, P.R. and McMahon, H.T. (2002) Curvature of clathrin-coated pits driven by epsin. Nature 419: 361-366 (abstract). (.pdf) supplemental data. (see also News and Views and highlights in Nature Cell Biology, Nature reviews in molecular cell biology, and a mini review in Cell (Cell October 18, 2002: 111 (2):143-146)(summary) ...
Thus was my situation this afternoon. A patient brought in this morning, down for hours at home following a cardiac arrest. Minimally responsive, with a poor prognosis, the attending had a conference with the family to explain these things and let them know that we would watch him for 72 hours to get a better idea of his neurological status and possibility of recovery. That afternoon, the attending left to place a difficult central line, and the patients oxygen dropped from 96 to 30, pressure dropped, pupils were dilated and nonreactive, no response to sternal rub- all signs of neurologic death and rapidly approaching cardiopulmonary death as well. Since he didnt have an end of life directive (at age 44, who would?), it needed to be ascertained quickly from his wife and daughter what their wishes were. And there was no other physician there. So, I put my big-girl coat on and was the physician, kneeling next to his wife, explaining his new signs and that a decision was needed more prematurely ...
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This meeting devoted to macropinocytosis will bring together experts from disparate fields with a shared interest in the biology of macropinosome formation and trafficking, with the goal of fostering collaboration and building a community.
Cellular processesCellular processesSporulation and germinationspore coat assembly protein SafA (TIGR02899; HMM-score: 63.9) ...
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Endocytosis is a mechanism for cells to remove ligands, nutrients, and plasma membrane (PM) proteins, and lipids from the cell surface, bringing them into the cell interior. Transmembrane proteins entering through clathrin-dependent endocytosis (CDE) have sequences in their cytoplasmic domains that bind to the APs (adaptor-related protein complexes) and enable their rapid removal from the PM. In addition to APs and clathrin, there are numerous accessory proteins including dynamin. Depending on the various proteins that enter the endosome membrane, these cargoes are sorted to distinct destinations. Some cargoes, such as nutrient receptors, are recycled back to the PM. Ubiquitylated membrane proteins, such as activated growth-factor receptors, are sorted into intraluminal vesicles and eventually end up in the lysosome lumen via multivesicular endosomes (MVEs). There are distinct mechanisms of clathrin-independent endocytosis (CIE) depending upon the cargo and the cell type ...
A leader among scientists, Sandra Schmid, Ph.D., was named Chair of the Department of Cell Biology in 2011. Dr. Schmid is committed to mentoring young scientists and gives career development and time management seminars to postdoctoral fellows and junior faculty nationwide. Her research focuses on clathrin-mediated endocytosis, the major pathway for uptake into the cell, with a particular emphasis on the GTPase dynamin. Dr. Schmid was co-founding editor of Traffic, Editor-in-Chief of Molecular Biology of the Cell, and is a past president of the American Society for Cell Biology (ASCB). Her awards include an ASCB/ WICB Junior Career Recognition Award, an NIH MERIT Award, and the William C. Rose Award from the American Society for Biochemistry and Molecular Biology. She is a Fellow of AAAS. Dr. Schmid earned her doctorate at Stanford University and recently earned a Master of Science in Executive Leadership at the University of San Diego. She came to UT Southwestern from The Scripps Research ...
PubMed Central Canada (PMC Canada) provides free access to a stable and permanent online digital archive of full-text, peer-reviewed health and life sciences research publications. It builds on PubMed Central (PMC), the U.S. National Institutes of Health (NIH) free digital archive of biomedical and life sciences journal literature and is a member of the broader PMC International (PMCI) network of e-repositories.
The central finding of the present study is that Mt3 plays a key role in the clathrin-dependent endocytosis of Aβ in astrocytes. In Mt3 −/− astrocytes, clathrin-mediated endocytosis, the mechanism responsible for Aβ endocytosis, was markedly decreased, whereas caveolin-mediated endocytosis was not altered. Astrocytes are likely key players in the clearance of extracellular Aβ; thus, our results suggest that changes in the Mt3 expression in astrocytes may have clinical relevance in AD. Taken together with our previous findings that Mt3 helps to maintain lysosomal degradation in astrocytes, the reduction in Mt3 in astrocytes may aggravate Aβ accumulation in the extracellular space.. Early studies showed that AD brain extracts induce more neurite outgrowth in cell cultures than do control brain extracts [27], suggesting upregulation of a growth-inducing factor or downregulation of a growth-inhibitory factor (GIF) in AD brains. The latter was shown to be the case, and a subsequent study ...
Nucleotide binding to the 70 kDa heat-shock cognate protein (Hsc70) from mung bean seeds and pig brain was investigated, as well as the clathrin uncoating activity of Hsc70 in the presence of these nucleotides. The two enzymes were found to behave identically. ATP bound to two different forms of Hsc70, with dissociation constants of 1.1±0.1 µM and 1.4±0.7 mM respectively at 25 °C. This corresponds to ΔG0´ = -34 and -16 kJ/mol respectively. From the temperature-dependence of the dissociation constant of the high-affinity site, ΔH0´ was calculated to -36±2 kJ/mol. This gives ΔS0´ = 6.7 J/mol per K. Adenosine 5´-[γ-thio]triphosphate, ADP, adenosine 5´-[β,γ-imino]triphosphate and adenosine 5´-[β,γ-methylene]triphosphate showed dissociation constants of 2.3, 11, 31 and 284 µM respectively. The order of affinities corresponded to the order of effectiveness in uncoating of pig brain coated vesicles. The implications of these findings for the mechanism of Hsc70 action are discussed. ...
Tor2 is an activator of the Rom2/Rho1 pathway that regulates α-factor internalization. Since the recruitment of endocytic proteins such as actin binding proteins and the amphiphysins precedes the internalization of α-factor, I hypothesized that loss of Tor function leads to an alteration in the dynamics of the endocytic proteins. I report here that endocytic proteins, Abp1 and Rvs167, are less recruited to endocytic sites not only in tor2 but also tor1 mutants. Furthermore, I found that the endocytic proteins Rvs167 and Sjl2 are completely mistargeted to the cytoplasm in tor1∆tor2ts double mutant cells. I also demonstrate here that the efficiency of endocytic internalization or scission in all tor mutants was drastically decreased. In agreement with the Sjl2 mislocalization, I found that in tor1∆tor2ts double mutant cells, as well as other tor mutant cells, the overall PIP2 level was dramatically increased. Finally, the cell wall chitin content in tor2ts and tor1∆tor2ts mutant cells was also
Angiogenesis: Growth of new blood vessels by sprouting from existing ones. Anoxia: a condition characterized by an absence of oxygen supply to an organ or a tissue Apoptosis: Form of cell death, also known as programmed cell death, in which a suicide program is activated within the cell, leading to fragmentation of the DNA, shrinkage of the cytoplasm, membrane changes and cell death without lysis or damage to neighboring cells. It is a normal phenomenon, occurring frequently in a multicellular organism. ARF: ADP Ribosylation Factor (ARF) is a member of the GTP-binding proteins responsible for regulating both COPI coat assembly and clathrin coat assembly at Golgi membranes. ATM: a protein that regulates several cellular responses to DNA breaks. C. elegans: Caenorhabditis elegans is a nematode (unsegmented) worm with very simple anatomy. Chaperone (molecular chaperone): Protein that helps other proteins avoid misfolding pathways that produce inactive or aggregated polypeptides. Drosophila: ...
TY - JOUR. T1 - Transforming growth factor β receptor signaling and endocytosis are linked through a COOH terminal activation motif in the type I receptor. AU - Garamszegi, N.. AU - Doré, Jr. AU - Penheiter, S. G.. AU - Edens, M.. AU - Yao, D.. AU - Leof, E. B.. PY - 2001/1/1. Y1 - 2001/1/1. N2 - Transforming growth factor β (TGF-β) coordinates a number of biological events important in normal and pathophysiological growth. In this study, deletion and substitution mutations were used to identify receptor motifs modulating TGF-β receptor activity. Initial experiments indicated that a COOH-terminal sequence between amino acids 482-491 in the kinase domain of the type I receptor was required for ligand-induced receptor signaling and down-regulation. These 10 amino acids are highly conserved in mammalian, Xenopus, and Drosophila type I receptors. Although mutation or deletion of the region (referred to as the NANDOR BOX, for nonactivating non-down-regulating) abolishes TGF-β-dependent ...
Looking for coated pit? Find out information about coated pit. A cell surface depression that is coated with clathrin on its cytoplasmic surface and functions in receptor-mediated endocytosis Explanation of coated pit
... interactions with clathrin, and the impact of overexpression on clathrin-mediated traffic". Mol. Biol. Cell. 10 (8): 2687-702. ... Goodman OB, Keen JH (1995). "The alpha chain of the AP-2 adaptor is a clathrin binding subunit". J. Biol. Chem. 270 (40): 23768 ... Kirchhausen T (2000). "Clathrin". Annu. Rev. Biochem. 69: 699-727. doi:10.1146/annurev.biochem.69.1.699. PMID 10966473. ... This gene encodes the alpha 1 adaptin subunit of the adaptor protein 2 (AP2 adaptors) complex found in clathrin coated vesicles ...
"Chromosome localization of human genes for clathrin adaptor polypeptides AP2 beta and AP50 and the clathrin-binding protein, ... Kalthoff C, Groos S, Kohl R, Mahrhold S, Ungewickell EJ (Nov 2002). "Clint: a novel clathrin-binding ENTH-domain protein at the ... Kirchhausen T (2000). "Clathrin". Annual Review of Biochemistry. 69: 699-727. doi:10.1146/annurev.biochem.69.1.699. PMID ... Kim YM, Benovic JL (Aug 2002). "Differential roles of arrestin-2 interaction with clathrin and adaptor protein 2 in G protein- ...
Kirchhausen T (2000). "Clathrin". Annu. Rev. Biochem. 69: 699-727. doi:10.1146/annurev.biochem.69.1.699. PMID 10966473. Geyer M ... 1999). "A dileucine motif in HIV-1 Nef acts as an internalization signal for CD4 downregulation and binds the AP-1 clathrin ... 1999). "A dileucine motif in HIV-1 Nef is essential for sorting into clathrin-coated pits and for downregulation of CD4". Curr ... Doray B, Kornfeld S (2001). "Gamma subunit of the AP-1 adaptor complex binds clathrin: implications for cooperative binding in ...
The protein encoded by this gene is part of the clathrin coat assembly complex which links clathrin to receptors in coated ... Kirchhausen T (2000). "Clathrin". Annual Review of Biochemistry. 69: 699-727. doi:10.1146/annurev.biochem.69.1.699. PMID ... Fölsch H, Ohno H, Bonifacino JS, Mellman I (Oct 1999). "A novel clathrin adaptor complex mediates basolateral targeting in ... Fölsch H, Ohno H, Bonifacino JS, Mellman I (Oct 1999). "A novel clathrin adaptor complex mediates basolateral targeting in ...
"Tandem arrangement of the clathrin and AP-2 binding domains in amphiphysin 1 and disruption of clathrin coat function by ... Kirchhausen T (2000). "Clathrin". Annual Review of Biochemistry. 69: 699-727. doi:10.1146/annurev.biochem.69.1.699. PMID ... "Crystal structure of the alpha appendage of AP-2 reveals a recruitment platform for clathrin-coat assembly". Proceedings of the ... "Association and colocalization of Eps15 with adaptor protein-2 and clathrin". The Journal of Cell Biology. 136 (4): 811-21. doi ...
Kirchhausen T (2000). "Clathrin". Annual Review of Biochemistry. 69: 699-727. doi:10.1146/annurev.biochem.69.1.699. PMID ... "A dileucine motif in HIV-1 Nef acts as an internalization signal for CD4 downregulation and binds the AP-1 clathrin adaptor". ... "A dileucine motif in HIV-1 Nef is essential for sorting into clathrin-coated pits and for downregulation of CD4". Current ... where it mediates both the recruitment of clathrin to the membrane and the recognition of sorting signals within the cytosolic ...
Budding of the plasma membrane then occurs, forming a clathrin coated pit.[1] Other receptors can nucleate a clathrin-coated ... Receptor-mediated endocytosis (RME), also called clathrin-mediated endocytosis, is a process by which cells absorb metabolites ... Clathrin-mediated endocytosis of many receptor types begins with the cargo ligands in the luminal compartment of the cell ... Kaksonen M, Roux A (May 2018). "Mechanisms of clathrin-mediated endocytosis". Nature Reviews. Molecular Cell Biology. 19 (5): ...
However, in higher eukaryotes there are several conserved motifs such as the clathrin-binding motifs which bind clathrin heavy ... The epsin homologue of C. elegans is EPN-1. EPN-1 conserves the UIM, ENTH domain, and clathrin-binding motif. ... Chen X, Irani NG, Firml, J (2011). "Clathrin - mediated endocytosis: the gateway into plant cells". Current Opinion in Plant ... Epsin 4, which encodes the protein enthoprotin, now known as clathrin interactor 1 (CLINT1), has been shown to be involved in ...
IFNAR1 and IFNAR2 can be internalized through endocytosis in response to agonism through clathrin-dependent and clathrin- ... Electron microscopy experiments show IFNAR receptors concentrated in clathrin-coated pits, and inhibition of clathrin-mediated ... Clathrin-dependent endocytosis[edit]. Following receptor agonism, the C-terminus of IFNAR is phosphorylated, followed by its ... in the presence of stimulation when the clathrin-dependent endocytosis pathway is inhibited with siRNA knockdown of clathrin or ...
Fölsch H, Pypaert M, Schu P, Mellman I (Feb 2001). "Distribution and function of AP-1 clathrin adaptor complexes in polarized ... Korolchuk VI, Banting G (Jun 2002). "CK2 and GAK/auxilin2 are major protein kinases in clathrin-coated vesicles". Traffic. 3 (6 ... Fölsch H, Ohno H, Bonifacino JS, Mellman I (Oct 1999). "A novel clathrin adaptor complex mediates basolateral targeting in ... Hirst J, Robinson MS (Aug 1998). "Clathrin and adaptors". Biochimica et Biophysica Acta. 1404 (1-2): 173-93. doi:10.1016/S0167- ...
Jung N, Haucke V (September 2007). "Clathrin-mediated endocytosis at synapses". Traffic. 8 (9): 1129-36. doi:10.1111/j.1600- ... Long-term depression enacts mechanisms to decrease AMPA receptor density in selected dendritic spines, dependent on clathrin ... Interactions from calcineurin activate dynamin GTPase activity, allowing the clathrin pit to excise itself from the cell ... consisting of a clathrin-coated pit underneath a section of AMPAR-containing plasma membrane and interacting proteins, is the ...
5) In the periactive zone the membrane proteins are sequestered and are endocytosed forming a clathrin coated vesicle. (6) The ... Jung Nadja; Haucke Volker (2007). "Clathrin-Mediated Endocytosis at Synapses". Traffic. 8 (9): 1129-1136. doi:10.1111/j.1600- ... 2010). "Intersectin 1 forms complexes with SGIP1 and Reps1 in clathrin-coated pits". Biochemical and Biophysical Research ... clathrin and endophilin.[17] In Drosophilia the intersectin homolog, Dap160, is located in the periactive zone of the ...
1997). "Arrestin/clathrin interaction. Localization of the arrestin binding locus to the clathrin terminal domain". J. Biol. ... 1998). "Clathrin-mediated endocytosis of the beta-adrenergic receptor is regulated by phosphorylation/dephosphorylation of beta ... ter Haar E, Musacchio A, Harrison SC, Kirchhausen T (1998). "Atomic structure of clathrin: a beta propeller terminal domain ... interaction of beta-arrestin with the AP-2 adaptor is required for the clustering of beta 2-adrenergic receptor into clathrin- ...
The holotoxin can be taken up by clathrin-coated pits, as well as by clathrin-independent pathways including caveolae and ... Nichols BJ, Lippincott-Schwartz J (October 2001). "Endocytosis without clathrin coats". Trends Cell Biol. 11 (10): 406-12. doi: ...
P2 and clathrin by AP180 in the nucleation of clathrin lattices on membranes". Science. 291 (5506): 1051-5. doi:10.1126/science ... Pearse, B. M. F.; Robinson, M. S. (1990). "Clathrin, Adaptors, and Sorting". Annual Review of Cell Biology. 6: 151-171. doi: ... Pearse first purified coated vesicles; she also discovered the clathrin coat molecule in 1975. Coated pits and vesicles were ... subscription required) Pearse, B. M. (1987). "Clathrin and coated vesicles". The EMBO Journal. 6 (9): 2507-12. PMC 553666 . ...
Byland R, Vance PJ, Hoxie JA, Marsh M (2007). "A Conserved Dileucine Motif Mediates Clathrin and AP-2-dependent Endocytosis of ... One of two major clathrin-associated adaptor complexes, AP-2, is a heterotetramer which is associated with the plasma membrane ... "Entrez Gene: AP2S1 adaptor-related protein complex 2, sigma 1 subunit". Pearse BM, Smith CJ, Owen DJ (2000). "Clathrin coat ... 2006). "Interaction of HIV-1 Gag with the clathrin-associated adaptor AP-2". Virology. 342 (2): 190-200. doi:10.1016/j.virol. ...
Examples include: Archain ARFs Clathrin Caveolin Dynamin and related proteins, such as the EHD protein family Rab proteins ... cis-Golgi Clathrin : trans-Golgi ----> Lysosomes, Plasma membrane ----> Endosomes (receptor-mediated endocytosis) Membrane ...
The clathrin coated pit invaginates into the cytosol and forms a clathrin coated vesicle. The clathrin proteins will then ... The particles are absorbed through the use of clathrin coated pits. These clathrin coated pits are short lived and serve only ... Proteins in the clathrin coat on the plasma membrane have propensity to bind and trap macromolecules or ligands. However, it is ... Clathrin Hydrolase Lysosome Phagocytosis Pinocytosis Rieger, R.; Michaelis, A.; Green, M.M. 1991. Glossary of Genetics. ...
Once the clathrin coat detaches, other proteins can interact directly with the AMPARs using PDZ carboxyl tail domains; for ... The complex, consisting of a clathrin-coated pit underneath a section of AMPAR-containing plasma membrane and interacting ... Long-term depression enacts mechanisms to decrease AMPA receptor density in selected dendritic spines, dependent on clathrin ... Interactions from calcineurin activate dynamin GTPase activity, allowing the clathrin pit to excise itself from the cell ...
The AP2 adaptor complex is a multimeric protein that works on the cell membrane to internalize cargo in clathrin-mediated ... The alpha and beta appendage domains bind to accessory proteins and to clathrin. Their interactions allow the temporal and ... "Clathrin coat construction in endocytosis". Current Opinion in Structural Biology. 10 (2): 220-8. doi:10.1016/S0959-440X(00) ... spatial regulation of the assembly of clathrin-coated vesicles and their endocytosis. The AP-2 complex is a heterotetramer ...
The CLINT1 protein binds to the terminal domain of the clathrin heavy chain and stimulates clathrin cage vesicle assembly. ... Clathrin interactor 1 (CLINT1), also known as EPSIN4, is a protein which in humans is encoded by the CLINT1 gene. ... Clathrin coated vesicles enable neurotransmitter receptors and other proteins to be endocytosed or taken up across neuronal ... 2004). "Clathrin adaptor epsinR is required for retrograde sorting on early endosomal membranes". Dev. Cell. 6 (4): 525-38. doi ...
The protein encoded by this gene is the medium chain of the trans-Golgi network clathrin-associated protein complex AP-1. The ... This complex is located at the Golgi vesicle and links clathrin to receptors in coated vesicles. These vesicles are involved in ... Foti M, Mangasarian A, Piguet V, Lew DP, Krause KH, Trono D, Carpentier JL (Oct 1997). "Nef-mediated clathrin-coated pit ... Fölsch H, Ohno H, Bonifacino JS, Mellman I (Oct 1999). "A novel clathrin adaptor complex mediates basolateral targeting in ...
"Dynamics of intracellular clathrin/AP1- and clathrin/AP3-containing carriers". Cell Reports. 2 (5): 1111-9. doi:10.1016/j. ... The involvement of the heterotetramer of COPI is similar to that of the AP/clathrin situation, but the coat of COPI is not ... Cocucci E, Aguet F, Boulant S, Kirchhausen T (August 2012). "The first five seconds in the life of a clathrin-coated pit". Cell ... AP-5 is associated with 2 proteins, SPG11 and SPG15, which have some structural similarity to clathrin, and may form the coat ...
Raiborg C, Bache KG, Mehlum A, Stang E, Stenmark H (2001). "Hrs recruits clathrin to early endosomes". EMBO J. 20 (17): 5008-21 ... "Hrs recruits clathrin to early endosomes". EMBO J. 20 (17): 5008-21. doi:10.1093/emboj/20.17.5008. PMC 125612 . PMID 11532964. ...
Cocucci E, Aguet F, Boulant S, Kirchhausen T (2012). "The first five seconds in the life of a clathrin-coated pit". Cell. 150 ( ... Henne WM, Boucrot E, Meinecke M, Evergren E, Vallis Y, Mittal R, McMahon HT (2010). "FCHo proteins are nucleators of clathrin- ... The muniscin protein family was initially defined in 2009 as proteins having 2 homologous domains that are involved in clathrin ... Robinson MS (2015). "Forty Years of Clathrin-coated Vesicles". Traffic. 16 (12): 1210-38. doi:10.1111/tra.12335. PMID 26403691 ...
May be required to determine the amount of membrane to be recycled, possibly by regulating the size of the clathrin cage. ... Involved in AP2-dependent clathrin-mediated endocytosis at the neuromuscular junction. Plays a crucial role in fetal and adult ... to cell membranes at sites of coated-pit formation and clathrin-vesicle assembly. ... Assembly protein recruiting clathrin and adapter protein complex 2 (AP2) ...
Clathrin-mediated endocytosis is the process by which cargo-containing clathrin-coated vesicles bud off from the plasma ... Simultaneous binding of PtdIns(4,5)P2 and clathrin by AP180 in the nucleation of clathrin lattices on membranes. Science. 291: ... Although the budding of clathrin-coated vesicles in vitro has recently been shown to only require the presence of clathrin, an ... However, these studies on PIP2 depletion reported different effects on clathrin assembly. In one study the number of clathrin ...
Clathrin-coated vesicles are the most prominent carriers of membrane traffic from cell surface to endosomes (endocytosis), a ... technological and analytical advances that I developed to directly visualize clathrin-mediated membrane traffic in three ... using these advances that defined a role for actin filament polymerization in counteracting membrane tension during clathrin- ...
In this study, the genes ZmCHC1 and ZmCHC2 encoding clathrin heavy chain in maize were cloned and characterized for the first ... a three-legged triskelion composed of three clathrin heavy chains (CHCs) and three light chains (CLCs), plays a critical role ... in clathrin-mediated endocytosis (CME) in eukaryotic cells. ... Clathrin, a three-legged triskelion composed of three clathrin ... Clathrin is a protein that plays a major role in the formation of coated vesicles. Clathrin was first isolated and named by ...
CB, clathrin-binding site; CB-I and CB-II: clathrin-binding enhancing sites I and II, respectively; EBD, endofin-binding domain ... Clathrin-binding assay. The clathrin-binding assay with GST fusion protein was adapted from the protocol described in a ... Optimal clathrin binding by TOM1 involves three sites: residues 300-321, 321-326 and a putative clathrin-binding box at ... The polyclonal anti-clathrin heavy chain antibody for western blotting and monoclonal anti-clathrin heavy chain for ...
Clathrin heavy chain (Chc) co-immunopurifies with Bic-D in embryos and ovaries, and a fraction of Chc colocalizes with Bic-D. ... Clathrin heavy chain plays multiple roles in polarizing the Drosophila oocyte downstream of Bic-D ... Clathrin heavy chain plays multiple roles in polarizing the Drosophila oocyte downstream of Bic-D ... Clathrin heavy chain plays multiple roles in polarizing the Drosophila oocyte downstream of Bic-D ...
In animal cells, dynamin and clathrin play central roles in vesicle formation in the process of endocytosis, but the roles of ... The localization pattern and behavior of the puncta were similar to those of animal dynamin and clathrin signals reported ... Imaging actin and dynamin recruitment during invagination of single clathrin-coated pits MERRIFIELD CJ ... Functional evidence for the identification of an Arabidopsis clathrin light chain polypeptide SCHEELE U ...
The function of clathrin is coordinated by clathrin adaptor proteins (Pearse and Robinson, 1984; Traub, 2003). The clathrin ... SMAP2 immunoprecipitated clathrin and AP-1 through a putative clathrin-binding domain and a CALM-binding domain, and SMAP2 ... Here, we found that clathrin, a vesicle coat protein, and clathrin adaptor protein complex 1 (AP-1) were present at recycling ... 2006). SMAP2, a novel ARF GTPase-activating protein, interacts with clathrin and clathrin assembly protein and functions on the ...
Assembly polypeptides from coated vesicles mediate reassembly of unique clathrin coats. Clathrin assembly protein AP-2 induces ... Here we show that the AP-2 adaptor, a clathrin-associated coat-protein complex that nucleates clathrin-coated vesicle formation ... AP-2-containing clathrin coats assemble on mature lysosomes. L M Traub, L M Traub ... L M Traub, S I Bannykh, J E Rodel, M Aridor, W E Balch, S Kornfeld; AP-2-containing clathrin coats assemble on mature lysosomes ...
Clathrin heavy chain \ 11-526-C025 for more molecular products just contact us ... Clathrin heavy chain on chromosome 17) (CLH-17). [PICALM CALM] Phosphatidylinositol-binding clathrin assembly protein (Clathrin ... Clathrin assembly protein complex 2 mu medium chain) (Clathrin coat assembly protein AP50) (Clathrin coat-associated protein ... Clathrin assembly protein complex 1 mu-1 medium chain 1) (Clathrin coat assembly protein AP47) (Clathrin coat-associated ...
The nuclear export signal within clathrin assembly lymphoid myeloid leukemia protein (CALM) is critical for in vitro ... fig02: The nuclear export signal within clathrin assembly lymphoid myeloid leukemia protein (CALM) is critical for in vitro ... fig02: The nuclear export signal within clathrin assembly lymphoid myeloid leukemia protein (CALM) is critical for in vitro ... is found in a variety of hematopoietic malignancies.Wild-type clathrin assembly lymphoid myeloid leukemia protein (CALM) ...
We show an effect on the G2/M phase population of clathrin knockdown in HEK293 cells but show that repressing clathrin ... Clathrin is a multimeric protein involved in vesicle coat assembly. Recently clathrin distribution was reported to change ... was developed in which the endogenous clathrin heavy chain alleles were replaced with the human clathrin heavy chain under the ... We conclude that the association of clathrin with the mitotic spindle and the contribution of clathrin to endocytosis are ...
Clathrin is a protein that plays a major role in the formation of coated vesicles. Clathrin was first isolated and named by ... This is how clathrin gets its name, from the Latin clathratus meaning like a lattice. Coat-proteins, like clathrin, are used to ... During mitosis, clathrin binds to the spindle apparatus, in complex with two other proteins: TACC3 and ch-TOG/CKAP5. Clathrin ... Model of Clathrin assembly). *. Pérez-Gómez J, Moore I (March 2007). "Plant endocytosis: it is clathrin after all". Current ...
Endocytosis without clathrin coats.. Nichols BJ1, Lippincott-Schwartz J.. Author information. 1. MRC Laboratory of Molecular ... Many markers for clathrin-independent endocytosis are found in detergent-resistant membrane fractions, or lipid rafts. We will ... Various non-clathrin endocytic mechanisms have been identified, including uptake through caveolae, macropinosomes and via a ... To date, most studies on endocytosis in mammalian cells have focused on pathways that start with uptake through clathrin-coated ...
If you know of any papers that use this antibody, please contact us at antibodies [at] alzforum [dot] org for consideration in the References section.. ...
Clathrin is a large heterohexameric protein complex composed of three heavy chains and three light chains. Clathrin molecules ... Clathrin-associated adaptor protein (AP) complexes are a stoichiometric coat component of CCVs alongside clathrin itself, and ... The connection of the clathrin scaffold to the membrane is mediated by clathrin adaptors, which can bind directly to both the ... clathrin. Clathrin assembles from three-legged individual components called triskelions to form a polygonal lattice around the ...
Aftiphilin contains a clathrin box, with two identified clathrin-binding motifs [PMID: 15811338, PMID: 15758025]. ... Aftiphilin is a component of the clathrin machinery in neurons.. FEBS Lett. 579 2177-84 2005 ...
Clathrin-mediated endocytosis persists during unperturbed mitosis.. Tacheva-Grigorova SK1, Santos AJ, Boucrot E, Kirchhausen T. ... The dynamics of clathrin-coated pit formation and the uptake of transferrin are maintained in naturally dividing cells but are ... B) Surface density of clathrin/AP2-coated structures (CCS/100 µm2) calculated from the number of AP2 spots at the top and ... C) Total number of clathrin/AP2-coated structures (CCS per cell) on the plasma membrane of a cell as determined by counting the ...
Thank you for your interest in spreading the word about Science.. NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.. ...
Immunogen = synthetic peptide: E E D P A A A F L A Q Q E S E I A G I E N D, corresp. to amino acids 23-44 of Cow Clathrin light chain. ...
Clathrin-independent carriers (CLICs) are prevalent tubulovesicular membranes responsible for non-clathrin mediated endocytic ...
Clathrin coats contain both clathrin (acts as a scaffold) and adaptor complexes that link clathrin to receptors in coated ... Clathrin adaptor proteins, also known as adaptins, are vesicular transport adaptor proteins associated with clathrin. These ... Adaptins recognise and bind to clathrin through their hinge region (clathrin box), and recruit accessory proteins that modulate ... One function of clathrin and AP2 complex-mediated endocytosis is to regulate the number of GABA(A) receptors available at the ...
Vassilopoulos et al. (see the Perspective by Orme and Bogan) now describe a function for CHC22 clathrin, a second isoform of ... A role for the CHC22 clathrin heavy-chain isoform in human glucose metabolism. Science 324, 1192-1196 (2009). [Abstract] [Full ... clathrin that is present in humans and not in mice. CHC22 participates in the biogenesis of the intracellular compartment that ...
reacts with clathrin heavy chain (CHC17); no cross-reaction with the muscle isoform CHC22. ...
This binding then stimulates clathrin lattice assembly. HIP1 is an obligate binding partner for Huntingtin, and loss of this ... Huntingtin-interacting protein 1, clathrin-binding domain (IPR032422). Short name: HIP1_clath-bd ... Clathrin light-chain binds to a flexible coiled-coil domain in HIP1 and induces a compact state that is refractory to actin ... It carries a highly conserved HADLLRKN sequence motif at its N terminus which effects the binding of HIP1R to clathrin light- ...
  • Receptor-mediated and fluid-phase endocytosis were significantly inhibited in this line following clathrin knockout, and we used this to explore the significance of clathrin heavy chain expression for cell cycle progression. (ox.ac.uk)
  • CONCLUSIONS/SIGNIFICANCE: We conclude that the association of clathrin with the mitotic spindle and the contribution of clathrin to endocytosis are evolutionarily conserved. (ox.ac.uk)
  • Clathrin coat formation on lysosomes is temperature dependent, displays an absolute requirement for ATP, and occurs in both semi-intact cells and on purified lysosomes, suggesting that clathrin-coated vesicles might regulate retrograde membrane traffic out of the lysosomal compartment. (rupress.org)
  • Consequently this work highlights the need for a more detailed molecular understanding of the recruitment and function of clathrin at the spindle, since the localisation but not the impact of clathrin on mitosis appears to be robust in plants, mammalian and chicken B-cells. (ox.ac.uk)
  • The nuclear export signal within clathrin assembly lymphoid myeloid leukemia protein (CALM) is critical for in vitro immortalization of cells by CALM-AF10. (nih.gov)
  • In other cell-lines silencing RNA has been used by others to knockdown clathrin expression resulting in an increase in the mitotic index of the cells. (ox.ac.uk)
  • We show an effect on the G2/M phase population of clathrin knockdown in HEK293 cells but show that repressing clathrin expression in the DKO-R cell-line has no effect on the size of this population. (ox.ac.uk)
  • METHODOLOGY/PRINCIPAL FINDINGS: Previously a chicken pre-B lymphoma cell line (DKO-R) was developed in which the endogenous clathrin heavy chain alleles were replaced with the human clathrin heavy chain under the control of a tetracycline-regulatable promoter. (ox.ac.uk)
  • Additionally, we showed that the ploidy and the recovery kinetics following cell cycle arrest with nocodazole were unchanged by repressing clathrin heavy chain expression. (ox.ac.uk)
  • Various non-clathrin endocytic mechanisms have been identified, including uptake through caveolae, macropinosomes and via a separate constitutive pathway. (nih.gov)
  • We show that XCL is endocytosed by the clathrin-dependent pathway, and is delivered to late endosome/lysosome compartments. (biomedsearch.com)
  • The results of this study for the first time demonstrate that not only the clathrin-dependent pathway but also macropinocytosis are involved in fish DNA enveloped virus entry, thus providing a convenient tactic for exploring the life cycle of DNA viruses. (sigmaaldrich.com)
  • Alphaviruses utilize the clathrin-mediated endocytic pathway. (wikipedia.org)
  • Researchers of the group of cellular and molecular neurobiology of the Bellvitge Biomedical Research Institute (IDIBELL) and the University of Barcelona, led by researcher Artur Llobet, have shown that synaptic levels of the protein clathrin are a determinant factor for synaptic plasticity of neurons. (eurekalert.org)
  • The encoded protein is part of the heterotetrameric AP-3 protein complex which interacts with the scaffolding protein clathrin. (wikipedia.org)
  • The clathrin heavy chain CHC-1 negatively regulates pulling forces acting on centrosomes during interphase and on spindle poles during mitosis in one-cell C. elegans embryos. (antibodies-online.com)
  • EGFR and PTEN localize to short-lived clathrin-coated pits (CCPs), and PTEN regulates CCP dynamics and signaling, suggesting compartmentalized signaling is crucial for proper signaling transduction. (biologists.org)
  • E) Upper panel: plot of the individual lifetimes of clathrin/AP2 pits from HeLa and BSC1 cells calculated from 4D time series obtained from the bottom and top surfaces during metaphase and cytokinesis, or from 2D and 4D time series from the bottom and top surfaces of interphase cells. (nih.gov)
  • A novel role for Rab5-GDI in ligand sequestration into clathrin-coated pits. (nih.gov)
  • We have used an in vitro assay for clathrin-coated pit assembly to identify a novel component required for the invagination of newly formed coated pits. (nih.gov)
  • Using immunocryoelectron microscopy, WNV particles were seen within clathrin-coated pits after 2 min postinfection. (asm.org)
  • We found that most capsids associated with fewer than five TfRs and that ∼25% of TfR-bound capsids laterally diffused into assembling clathrin-coated pits less than 30 s after attachment. (asm.org)
  • Our results indicate that a key evolutionary conserved function of epsin, in addition to other roles that include, as we show here, a low affinity interaction with SNAREs, is to help generate the force that leads to invagination and then fission of clathrin-coated pits. (elifesciences.org)
  • The deep invagination of clathrin-coated pits that leads to fission is assisted by actin, a protein that assembles into filaments that are suggested to generate the forces needed for this process. (elifesciences.org)
  • Overall, these results suggest that a main role of epsin is to help actin interact with the clathrin-coated pits and generate the force required for a pit to develop into a vesicle. (elifesciences.org)
  • The particles are absorbed through the use of clathrin coated pits. (wikipedia.org)
  • These clathrin coated pits are short lived and serve only to form a vesicle for transfer of particles to the lysosome. (wikipedia.org)
  • In the presence of AP180, clathrin lattices formed on the monolayer. (rcsb.org)
  • Clathrin assembly lymphoid myeloid leukemia protein (CALM) is a homologue of AP180 whose ANTH domain shares 81% sequence homology. (cam.ac.uk)
  • In this AP180 + clathrin experiment the ratio of clathrin:AP180 in the liposome pellet (+ tubes) is much greater than in the supernatant. (cam.ac.uk)
  • After dissolving the liposomes with Tx-100, polymerised clathrin remains assembled while AP180 and AP2 move back to the supernatant. (cam.ac.uk)
  • Inhibition of several kinases selectively affected CME in cancer cells, including inhibition of ERK1/2, which selectively inhibited CME by decreasing the rate of clathrin-coated pit (CCP) initiation. (pnas.org)
  • Aftiphilin contains a clathrin box, with two identified clathrin-binding motifs [ PMID: 15811338 , PMID: 15758025 ]. (ebi.ac.uk)
  • The Hsc70 molecular chaperone effects the uncoating reaction, and is guided to appropriate locations on clathrin lattices by the J-domain-containing co-chaperone molecule auxilin. (nih.gov)
  • The pit grows deeper over time as more clathrin molecules assemble, eventually resulting in a deeply invaginated clathrin-coated pit that encloses the cargo to be taken up by the cell. (elifesciences.org)
  • This family member may play a significant role in cargo molecules regulation and clathrin-coated vesicle assembly. (wikipedia.org)
  • B) Surface density of clathrin/AP2-coated structures (CCS/100 µm 2 ) calculated from the number of AP2 spots at the top and bottom surfaces of HeLa and BSC1 cells imaged during interphase, metaphase, and cytokinesis prior to abscission. (nih.gov)
  • Here, we have used electron cryomicroscopy to determine 12-A-resolution structures of in-vitro-assembled clathrin coats in association with a carboxy-terminal fragment of auxilin that contains both the clathrin-binding region and the J domain. (nih.gov)
  • The result was an increase in large clathrin-coated structures (CCS) on the surface of the cells. (fredhutch.org)
  • The clathrin structures that formed in these manipulated cells recruited integrin β1 normally. (fredhutch.org)
  • Although most of the clathrin structures were found to contain both Dab2 and AP2, the authors showed that integrin β1 and TfnR do not co-localize on the cell surface. (fredhutch.org)
  • In contrast, when a truncated HIP1 protein containing both the DPF-like motifs and the coiled-coil domain was overexpressed, large perinuclear vesicle-like structures containing HIP1, huntingtin, clathrin and endocytosed transferrin were observed, indicating that HIP1 is an endocytic protein, the structural integrity of which is crucial for maintenance of normal vesicle size in vivo. (nih.gov)
  • Edeling MA, Smith C, Owen D. Life of a clathrin coat: insights from clathrin and AP structures . (humpath.com)
  • The general objective of our lab is to understand the functions of clathrin-coated structures (CCSs) during the different steps of cancer development. (ipbs.fr)
  • Indeed, a chimeric protein consisting of TOM1 fused to two FYVE domains derived from endofin has the ability to recruit clathrin onto endosomal structures. (biologists.org)
  • Notably, perturbation of N-WASP-CK2 complex function showed that N-WASP controls the presence of F-actin at clathrin-coated structures. (biologists.org)
  • Title: Role of phosphatidylinositol clathrin assembly lymphoid-myeloid leukemia (PICALM) in intracellular amyloid precursor protein (APP) processing and amyloid plaque pathogenesis. (nih.gov)
  • Knockdown of clathrin or the clathrin adaptor AP-1 in HCVcc-infected human hepatoma cell cultures impaired viral secretion without altering intracellular HCVcc levels or apolipoprotein B (apoB) and apoE exocytosis. (asm.org)
  • To investigate the intracellular role of the clathrin heavy chain in living cells, we have used "antisense" RNA to engineer mutant Dictyostelium discoideum cells that are severely deficient in clathrin heavy chain expression. (rupress.org)
  • The main clathrin heavy chain, located on chromosome 17 in humans, is found in all cells. (wikipedia.org)
  • Clathrin heavy chain is often described as a leg, with subdomains, representing the foot (the N-terminal domain), followed by the ankle, distal leg, knee, proximal leg, and trimerization domains. (wikipedia.org)
  • This was originally determined from the structure of the proximal leg domain that identified and is composed of a smaller structural module referred to as clathrin heavy chain repeat motifs. (wikipedia.org)
  • Endogenous clathrin was detected using X22 mAb (anticlathrin heavy chain) shown in red. (nih.gov)
  • BHK cells with inducible expression of antisense to clathrin heavy chain was preincubated with methyl-β-cyclodextrin (10 mM) for 30 min. (nih.gov)
  • Clathrin light and heavy chain interface: alpha-helix binding superhelix loops via critical tryptophans. (nih.gov)
  • LC binding to clathrin heavy chain (HC) was characterized by genetic and structural approaches. (nih.gov)
  • Auxilin binding produces a local change in heavy-chain contacts, creating a detectable global distortion of the clathrin coat. (nih.gov)
  • Epidermal growth factor (EGF) binding to its receptor causes rapid phosphorylation of the clathrin heavy chain at tyrosine 1477, which lies in a domain controlling clathrin assembly. (nih.gov)
  • CHCR repeats in the clathrin heavy chain, Saccharomyces cerevisiae Vamp2 and human Vamp6 have been implicated in homooligomerization, suggesting that this may be the primary function of this repeat. (embl.de)
  • In this study, the genes ZmCHC1 and ZmCHC2 encoding clathrin heavy chain in maize were cloned and characterized for the first time in monocots. (mdpi.com)
  • To reveal the biological consequence of endofin-mediated endosomal recruitment of TOM1, we have identified the clathrin heavy chain as a major interacting protein for TOM1. (biologists.org)
  • Of importance, the interaction between clathrin light and heavy chain is very stable, such that the molecular dynamics of LCb-RFP faithfully represents the behavior of clathrin triskelia subunits (Hoffmann et al. (nih.gov)
  • Here, we have tested this hypothesis by replacing endogenous clathrin heavy chain in human cells with a panel of clathrin constructs. (biologists.org)
  • A single CHC molecule consists of an N-terminal seven-bladed β-propeller, a linker region, eight clathrin heavy chain repeat (CHCR0-7) segments, a proximal hairpin, a tripod region that is thought to be responsible for trimerisation, and a variable C-terminal segment (residues 1631-1675). (biologists.org)
  • In the present study, we aimed to test these two hypotheses by replacing endogenous clathrin heavy chain (CHC) in human cells with a variety of CHC constructs. (biologists.org)
  • Clathrin heavy chain is required for pinocytosis, the presence of large vacuoles, and development in Dictyostelium. (rupress.org)
  • Similarly, Northern blots showed an absence of clathrin heavy chain mRNA. (rupress.org)
  • Clathrin heavy chain-deficient Dictyostelium cells were viable, but exhibited growth rates twofold slower than parental cells. (rupress.org)
  • We interfered with clathrin heavy chain (CHC) function through mutants and dominant-negative approaches in Arabidopsis thaliana and established tools to manipulate clathrin function in a cell type-specific manner. (ugent.be)
  • However, in higher eukaryotes there are several conserved motifs such as the clathrin-binding motifs which bind clathrin heavy chain, these motifs flank a cluster of up to eight DP repeats which bind to AP2. (wikipedia.org)
  • Based on circular dichroism data, mapping and mutagenesis, LCa and LCb were represented as alpha-helices, aligned along the HC and, using molecular dynamics, a structural model of their interaction was generated with novel implications for LC control of clathrin assembly. (nih.gov)
  • We have identified Rab5 as a critical cytosolic component required for clathrin-coated pit function. (nih.gov)
  • Clathrin fluorescence displayed gradual but sustained recovery, reaching 50% after ∼115 s (Figure 4 and Figure 4 Video 1), demonstrating that subunits of the FCL basketwork are being exchanged with the cytosolic pool. (nih.gov)
  • Auxilin depletion causes self-assembly of clathrin into membraneless cages in vivo. (biomedsearch.com)
  • We also revealed a nonendocytic role for clathrin required for extracellular EPEC infections. (nih.gov)
  • Together, these data demonstrate a fundamental role for clathrin function in cell polarity, growth, patterning, and organogenesis in plants. (ugent.be)
  • It carries a highly conserved HADLLRKN sequence motif at its N terminus which effects the binding of HIP1R to clathrin light-chain EED regulatory site. (ebi.ac.uk)
  • This adaptor protein is known to contain a clathrin-binding motif and a motif that recognizes Dab2 cargoes like integrin β1. (fredhutch.org)
  • Although residues 321-326 could function independently as a weak clathrin-binding motif, deletion of amino acids 300-321 or mutation of 362 Leu and 364 Asp to Ala residues reduced the binding of clathrin to TOM1. (biologists.org)
  • EPN-1 conserves the UIM, ENTH domain, and clathrin-binding motif. (wikipedia.org)