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 kinetics of CHC phosphorylation in activated T cells was slow compared with the total protein tyrosine phosphorylation in cell lysates (not depicted). This delay suggested that other signaling events are initiated before CHC phosphorylation. We had previously found that the activation of c-Src kinase or Lyn kinase, a Src kinase family member, was required for CHC phosphorylation after EGFR and BCR stimulation, respectively (20, 21). To address whether the activity of a Src family kinase was necessary for CHC phosphorylation in activated T cells, we treated Jurkat cells with various concentrations of the Src family kinase inhibitor PP1 before stimulation with soluble anti-CD3 Ab. At increasing concentrations of PP1, but not the serine/threonine kinase inhibitor H7, the level of inducible CHC phosphorylation was diminished (Fig. 2 A). Additionally, in the presence of PP1, the basal level of CHC phosphorylation in Jurkat cells before the induction of TCR internalization was also decreased, ...

In all organisms, cell polarity is fundamental for most aspects of cell physiology. In many species and cell types, it is controlled by the evolutionarily conserved PAR-3, PAR-6 and aPKC proteins, which are asymmetrically localized at the cell cortex where they define specific domains. While PAR proteins define the antero-posterior axis of the early C. elegans embryo, the mechanism controlling their asymmetric localization is not fully understood. Here we studied the role of endocytic regulators in embryonic polarization and asymmetric division. We found that depleting the early endosome regulator RAB-5 results in polarity-related phenotypes in the early embryo. Using Total Internal Reflection Fluorescence (TIRF) microscopy, we observed that PAR-6 is localized at the cell cortex in highly dynamic puncta and depleting RAB-5 decreased PAR-6 cortical dynamics during the polarity maintenance phase. Depletion of RAB-5 also increased PAR-6 association with clathrin heavy chain (CHC-1) and this ...

Clathrin-mediated endocytosis is exploited by bacterial and viral pathogens during internalization. Humphries and Way review recent studies which highlight the fact that, in addition to a structural role, clathrin can function as a signalling platform during pathogen entry, and other studies revealing that, in conjunction with actin, clathrin is involved in pathogen cell-cell spread and release. The role of clathrin in pathogen entry has received much attention and has highlighted the adaptability of clathrin during internalization. Recent studies have now uncovered additional roles for clathrin and have

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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 ...

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.. ...

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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 ...

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ALK a tyrosine kinase of the ALK family. Plays an important role in the development of the brain and exerts its effects on specific neurons in the nervous system. Translocated and expressed as a fusion protein in anaplastic lymphoma. About one third of large-cell lymphomas are caused by a t(2;5)(p23;q35) translocation that fuses ALK to nucleophosmin (NPM1A). Other cases caused by fusions of ALK to moesin, non-muscle myosin heavy chain 9, clathrin heavy chain and other genes. Several fusions also seen in inflammatory myofibroblastic tumors, and expression has been briefly noted in a range of tumors Note: This description may include information from UniProtKB ...

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. ...

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Eps15 is a substrate of the EGF receptor, and exhibits homology with the yeast protein End3, a protein involved in the endocytosis of the α factor in S. cerevisiae (Fazioli et al., 1993; Benedetti et al., 1994). Recently, Eps15 has been shown to bind directly to the ear of α-adaptin, a component of the AP-2 complex (Benmerah et al., 1995). In this paper, we have investigated the possible role of Eps15 in EGF receptor endocytosis. We have shown that Eps15 phosphorylation is mediated by the EGF receptor, and not by PDGF or insulin receptors, which suggest a possible function of Eps15 specifically in the endocytosis of EGF receptors. Immunoprecipitation studies revealed that Eps15 can be co-immunoprecipitated with the EGF receptor. Moreover, the association of Eps15 with the EGF receptor increased dramatically after activation of the receptor, while the association of Eps15 with AP-2 or clathrin LC appeared to occur independently of EGF. The association of Eps15 to the EGF receptor has not been ...

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

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 ...

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.

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.

CHC22 clathrin plays a key role in intracellular membrane traffic of the insulin-responsive glucose transporter GLUT4 in humans. We performed population genetic and phylogenetic analyses of the CHC22-encoding CLTCL1 gene, revealing independent gene loss in at least two vertebrate lineages, after arising from gene duplication. All vertebrates retained the paralogous CLTC gene encoding CHC17 clathrin, which mediates endocytosis. For vertebrates retaining CLTCL1 , strong evidence for purifying selection supports CHC22 functionality. All human populations maintained two high frequency CLTCL1 allelic variants, encoding either methionine or valine at position 1316. Functional studies indicated that CHC22-V1316, which is more frequent in farming populations than in hunter-gatherers, has different cellular dynamics than M1316-CHC22 and is less effective at controlling GLUT4 membrane traffic, attenuating its insulin-regulated response. These analyses suggest that ancestral human dietary change influenced ...

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 ...

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

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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.. ...

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.

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 ...

Thermodynamics of protein-mediated membrane deformation â application to clathrin dependent and clathrin independent endocytosis (2009 ...

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DI-fusion, le Dépôt institutionnel numérique de lULB, est loutil de référencementde la production scientifique de lULB.Linterface de recherche DI-fusion permet de consulter les publications des chercheurs de lULB et les thèses qui y ont été défendues.

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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

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. ...

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.

The nuclear export signal within clathrin assembly lymphoid myeloid leukemia protein (CALM) is critical for in vitro immortalization of cells by CALM-AF10. (a)

set by micropipette aspiration to be less than 1 over a wide range of spontaneous curvatures, indicating a high membrane-tension regime in their set up. Thus, our model is consistent with their observations of shallow buds observed in isotonic conditions. One result that our model cannot explain is the lack of any clathrin assembly observed under hypotonic conditions. It is possible that at extremely high membrane tensions, the coat is simply unable to stay bound to the membrane at the extremely flat morphology that would be expected.. Avinoam et al. (46) found that the size of the clathrin coat does not change substantially during membrane deformation in CME in human skin melanoma (SK-MEL-2) cells. This observation is in contrast to the canonical view that the clathrin coat should directly impose its preferred curvature on the underlying membrane (8). There are two possible explanations for this observation in the context of our study. One is that the membrane tension is too high for the coat ...

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 ...

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.. ...

Reactome is pathway database which provides intuitive bioinformatics tools for the visualisation, interpretation and analysis of pathway knowledge.

Reactome is pathway database which provides intuitive bioinformatics tools for the visualisation, interpretation and analysis of pathway knowledge.

In a prior post I documented the global impact of the CHC theory of intelligence. Another indicator of the impact of the CHC model and taxonomy is how it has been recognized and used forimportant research functions in psychology, other disciplines, and applied settings. A number of diverse examples are summarized here.The common CHC nomenclature assists different researchers better understan...

Control of E-cadherin apical localisation and morphogenesis by a SOAP-1/AP-1/clathrin pathway in C. elegans epidermal cells. ...

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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