Here, we isolated a novel F-actin-binding protein with a molecular mass of ∼205 kD (p205). This protein was copurified with another protein with a molecular mass of 190 kD (p190) that lacked the F-actin-binding activity on various column chromatographies. The molecular cloning of the cDNAs of these two proteins revealed that the nucleotide sequence of the p190 cDNA was identical to that of the p205 cDNA, except for the two splicing regions. FISH analysis revealed that the genes of these two proteins were localized at the same locus. These results suggest that p205 and p190 are splicing variants derived from the same gene. Because a computer homology search revealed that the aa sequence of p190 was almost identical to that of human AF-6 protein, we theorize that p190 may be a rat counterpart of human AF-6 protein. We named p205 and p190 l- and s-afadins, respectively. Further purification steps of l-afadin, including Mono S column and Superdex 200 column chromatographies, did not separate l- ...
TY - JOUR. T1 - Ezrin/radixin/moesin proteins differentially regulate endothelial hyperpermeability after thrombin. AU - Adyshev, Djanybek M.. AU - Dudek, Steven M.. AU - Moldobaeva, Nurgul. AU - Kim, Kyung mi. AU - Ma, Shwu Fan. AU - Kasa, Anita. AU - Garcia, Joe G.N.. AU - Verin, Alexander Dmitriyevich. PY - 2013/8/1. Y1 - 2013/8/1. N2 - Endothelial cell (EC) barrier disruption induced by inflammatory agonists such as thrombin leads to potentially lethal physiological dysfunction such as alveolar flooding, hypoxemia, and pulmonary edema. Thrombin stimulates paracellular gap and F-actin stress fiber formation, triggers actomyosin contraction, and alters EC permeability through multiple mechanisms that include protein kinase C (PKC) activation. We previously have shown that the ezrin, radixin, and moesin (ERM) actin-binding proteins differentially participate in sphingosine-1 phosphate-induced EC barrier enhancement. Phosphorylation of a conserved threonine residue in the COOH-terminus of ERM ...
TY - JOUR. T1 - Smooth muscle calponin. T2 - An unconventional CArG-dependent gene that antagonizes neointimal formation. AU - Long, Xiaochun. AU - Slivano, Orazio J.. AU - Cowan, Sarah L.. AU - Georger, Mary A.. AU - Lee, Ting Hein. AU - Miano, Joseph M.. PY - 2011/10/1. Y1 - 2011/10/1. N2 - OBJECTIVE-: Smooth muscle calponin (CNN1) contains multiple conserved intronic CArG elements that bind serum response factor and display enhancer activity in vitro. The objectives here were to evaluate these CArG elements for activity in transgenic mice and determine the effect of human CNN1 on injury-induced vascular remodeling. METHODS AND RESULTS-: Mice carrying a lacZ reporter under control of intronic CArG elements in the human CNN1 gene failed to show smooth muscle cell (SMC)-restricted activity. However, deletion of the orthologous sequences in mice abolished endogenous Cnn1 promoter activity, suggesting their necessity for in vivo Cnn1 expression. Mice carrying a 38-kb bacterial artificial ...
Cell adhesion to extracellular matrix is an important physiological stimulus for organization of the actin-based cytoskeleton. Adhesion to the matrix glycoprotein Thrombospondin-1 triggers the sustained formation of F-Actin microspikes that contain the Actin-bundling protein Fascin. These structures are also implicated in cell migration, which may be an important function of Thrombospondin-1 in tissue remodeling and wound repair. Fascin bundles Actin microfilaments within dynamic cellular structures such as microspikes, stress fibers and membrane ruffles. It may serve as a prognostic factor for abnormal ovarian epithelial pathology and may be a novel target for the treatment of ovarian cancer. An Actin-bundling protein, Fascin identifies dendritic cells in the blood and in tissues.. Clone: ...
Tumor suppressor which has the ability to inhibit cell growth and be pro-apoptotic during cell stress. Inhibits cell growth in bladder and prostate cancer cells by a down-regulation of HSPB1 by inhibiting its phosphorylation. May act as a capping or branching protein for keratin filaments in the cell periphery. May regulate K8/K18 filament and desmosome organization mainly at the apical or peripheral regions of simple epithelial cells (PubMed:15731013, PubMed:18931701). Is a negative regulator of ciliogenesis (PubMed:25270598 ...
Calponin 2 is a protein that in humans is encoded by the CNN2 gene. The CNN2 gene is located at 19p13.3 in the human chromosomal genome, encoding the protein calponin 2. Calponin 2 is one of the three isoforms of calponin and an actin filament-associated regulatory protein with wide tissue distributions. Human calponin 2 is a 33.7-kDa protein consisting of 309 amino acids with an isoelectric point (pI) of 7.23. Accordingly, it is also known as neutral calponin. Calponin isoforms are conserved proteins whereas calponin 2 has diverged from calponin 1 and calponin 3 mainly in the C-terminal variable region. Phylogenetic lineage of calponin 2 showed that calponin 2 is conserved among mammalian species but more diverged among amphibian, reptile and fish species (Fig 1). CNN2 is expressed in a broader range of tissue and cell types, including developing and remodeling smooth muscle as well as adult mature smooth muscle, epidermal keratinocytes, fibroblasts, lung alveolar cells, endothelial cells, ...
In the present study, we describe the cloning and functional characterization of the actin-binding protein myopodin, a novel member of the synaptopodin gene family (Mundel et al., 1997a). Myopodin and synaptopodin share a high degree of homology at the primary sequence level as well as some fundamental structural and functional properties. On the other hand, they also show significant differences that may allow them to exert different functions in the cell.. As described previously for synaptopodin (Mundel et al., 1997a), myopodin seems to be differentially modified in different tissues. Northern blot analysis revealed only one message of ∼3.6 kb in both heart and skeletal muscle (Fig. 2 a). However, by Western blot analysis, a protein of ∼80 kD was detected in skeletal muscle, whereas the corresponding protein in the heart had a size of ∼95 kD (Fig. 2 b). The cause of this size difference remains to be established, but for several reasons, it appears to be due to posttranslational ...
SM22α is a shape change and transformation sensitive 22 kDa actin-binding protein of the calponin family. It is ubiquitous to vascular and visceral smooth muscle, and is an early marker of smooth muscle differentiation. It is also present in fibrob
The major cause of death in pancreatic cancer is due to metastases; therefore, it is important to study the mechanism by which the pancreatic cancer cells migrate and invade. This would help advance therapeutics and ultimately help prolong survival. Adenylyl cyclase-associated protein 1 (CAP1) is a scaffold protein that is involved in the regulation of actin microfilament formation, which ultimately leads to cell migration and invasion. CAP1 binds to G-actin inhibiting polymerization. We first tested whether CAP1 binds to adenylyl cyclase (AC) by performing co-immunoprecipitation. We found that CAP1 not only interacts with G-actin, but also with a number of AC isoforms: AC1, AC3, AC4 and AC7. Further studies need to be done to determine how CAP1/AC/G-actin interact and the impact of these interactions on the invasive behavior of pancreatic cancer cells ...
NMR structural characterization of the N-terminal domain of the adenylyl cyclase-associated protein (CAP) from Dictyostelium discoideum. Related
The 65-kDa protein (p65) was previously identified as a phosphorylated protein in activated macrophages, and has turned out to be a member of a plastin protein family characterized by a series of Ca,SUP,2+,/SUP,-, calmodulin-, and β-actin-binding domains. In mice, two isoforms, p65/L-plastin and T-plastin, have so far been identified; p65/L-plastin is expressed in hemopoietic cells and cancer cells, and T-plastin in solid tissue cells. We generated monoclonal antibodies to p65/L-plastin, examined the isoform-specificity by using recombinant (r) T-plastin, and found that the antibodies were specific for rp65/L-plastin, whereas immune sera to rp65/L-plastin showed cross-reactions to rT-plastin. One of the antibodies, p65-7B5, was demonstrated to react to native p65/L-plastin by Western blot, flow cytometric, and immunohistochemical analysis. Furthermore, p65-7B5 has made it possible to detect p65/L-plastin-expressing cells in tissues where T-plastin is abundantly expressed. These reagents and ...
Human fibrillin-1 is the major structural protein of extracellular matrix 10-12 nm microfibrils. It has a disulfide-rich modular organization which consists primarily of cbEGF (Ca 2+ -binding epidermal growth factor-like) domains and TB (transforming growth factor β-binding protein-like) domains. TB4 contains an RGD (Arg-Gly-Asp) integrin-binding motif. The atomic structure of this region has been solved by X-ray crystallography and shows the TB4 and flanking cbEGF domains to be arranged as a tetragonal pyramid with N- and C-termini exposed at opposite ends of the fragment. The RGD integrin-binding motif is located within a flexible loop. We have used a variety of biophysical, biochemical and cell biology methods to investigate the molecular properties of integrin-fibrillin-1 interactions and have demonstrated that recombinant fibrillin-1 domain fragments mediate binding to integrins αVβ3, α5β1 and αVβ6. Integrin αVβ3 is a high-affinity fibrillin-1 receptor (K d ∼40 nM), whereas integrins
Fascin is a actin cross-linking protein.The Fascin gene contains 5 exons and spans 7 kb. It is a 54-58 kilodalton monomeric actin filament bundling…
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Moesin is a member of the ezrin-radixin-moesin (ERM) family of proteins that are important for organizing membrane domains and receptor signaling and regulating the migration of effector T cells. Whether moesin plays any role during the generation of TGF-β-induced Tregs (iTregs) is unknown. Here, we have discovered that moesin is translationally regulated by TGF-β and is also required for optimal TGF-β signaling that promotes efficient development of iTregs. Loss of moesin impaired the development and function of both peripherally derived iTregs and in vitro-induced Tregs. Mechanistically, we identified an interaction between moesin and TGF-β receptor II (TβRII) that allows moesin to control the surface abundance and stability of TβRI and TβRII. We also found that moesin is required for iTreg conversion in the tumor microenvironment, and the deletion of moesin from recipient mice supported the rapid expansion of adoptively transferred CD8+ T cells against melanoma. Our study establishes ...
Fascin 1 is an actin-bundling protein that is dramatically overexpressed in a variety of invasive tumors and thought to have a critical role in cancer cell metastasis. However, as a drug target it is highly challenging due to its mechanism of protein-protein interaction and the lack of knowledge around the critical actin-binding sites. Using a fragment-based approach, biophysical assay screening and X-ray crystallography, we have been able to identify and optimize novel fascin 1 inhibitors. Furthermore we have developed robust and reproducible biochemical binding and bundling assays which have allowed us to develop ligands with submicromolar affinity.. Fascin 1 cross-links filamentous actin (F-actin) into parallel bundles that are involved in the formation of dynamic cellular protrusions (such as lamellipodia and filopodia) used during cell migration. It also contributes to the formation of actin-rich finger-like protrusions, termed invadopodia, that tumor cell lines use to degrade the tumor ...
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During animal development cells assemble into tissues and establish specific adhesion sites with the surrounding extracellular matrix (ECM). ECM enables the stable attachment of cells and their separation in distinct layers permitting tissue morphogenesis. The integrin family of transmembrane proteins mediate physical contact of cells with the ECM. This connection requires direct binding of the extracellular domain of both the α- and the β-subunit of integrin to their ECM ligands, whereas their small cytoplasmic tails bind a network of proteins to form the integrin adhesome and mediate the link to the actin cytoskeleton (Geiger and Yamada, 2011). The molecular composition of the integrin adhesome network is diverse, highly dynamic and largely determined by the cell type and the physical strength required by the local developmental microenvironment (Wolfenson et al., 2009; Zaidel-Bar et al., 2007).. The tripartite IPP complex containing integrin-linked kinase (ILK), PINCH and parvin is central ...
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Patients with triple-negative breast cancer (TNBC) have an overall poor prognosis, which is primarily due to a high metastatic capacity of these tumors. Novel therapeutic approaches to target the signaling pathways that promote metastasis are desirable, in order to improve the outcome of TNBC patients. A loss of function of a microRNA, miR-206, is related to increased metastasis potential in breast cancers but the mechanism remains to be elucidated. In this study, we show that miR-206 was decreased in TNBC clinical tumor samples and cell lines whereas one of its predicted targets, actin-binding protein CORO1C, was increased. Expression of miR-206 significantly reduced proliferation by inducing a G1-S cell cycle arrest and migration and also repressed CORO1C mRNA and protein levels. We demonstrate that miR-206 interacts with the 3-untranslated region (3-UTR) of CORO1C and regulates this gene post-transcriptionally. Further, silencing of CORO1C reduced tumor cell migration and affected the actin ...
May be a crucial component of the cytoskeleton of highly motile cells, functioning both in the invagination of large pieces of plasma membrane, as well as in forming protrusions of the plasma membrane involved in cell locomotion.
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mouse Dbnl protein: a 55 kDa actin-binding cytoskeleton adapter protein that is coupled to signal transduction from lymphocyte antigen receptors; a substrate for Src & Syk kinases; isolated from mouse; RefSeq NM_013810
1HQZ: Phased translation function revisited: structure solution of the cofilin-homology domain from yeast actin-binding protein 1 using six-dimensional searches.
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The only ethnic supremacists in these fora for Bolaji Aluko are people like Obi Nwakanma and Nebu - so says the uber ethnic supremacist himself by the name, Bolaji Aluko. I mentioned the fact that Chima is Igbo simply because Abba insisted that Southerners should not presume to know the problems in the North or proffer solutions to what they know nothing about. Professor Chimas work was dedicated to nomadic education, particularly the education of the pastoralist communities in the North, and he did not have to be a Fulani to know the problem or the solution to the crisis of education in the North which Abba denies. For mentioning Chimas Igbo identity, Obi Nwakanma becomes supremacist - and largely because the word Igbo raises the hackles of the likes of Bolaji Aluko. The problem is simple: the crisis of education in the North is a Nigerian problem, and requires all hands on deck: it does not matter from where the solutions come, - northerner or Southerner. If that makes me ...
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MEGTHCTLQLHKPITELCYISFCLPKGEVRGFSYKGTVTLDRSNKGFHNCYQVREESDIISLSQEPDEHP 1 - 70 GDIFFKQTPTKDILTELYKLTTERERLLTNLLSSDHILGITMGNQEGKLQELSVSLAPEDDCFQSAGDWQ 71 - 140 GELPVGPLNKRSTHGNKKPRRSSGRRESFGALPQKRTKRKGRGGRESAPLMGKDKICSSHSLPLSRTRPN 141 - 210 LWVLEEKGNLLPNGALACSLQRRESCPPDIPKTPDTDLGFGSFETAFKDTGLGREVLPPDCSSTEAGGDG 211 - 280 IRRPPSGLEHQQTGLSESHQDPEKHPEAEKDEMEKPAKRTCKQKPVSKVVAKVQDLSSQVQRVVKTHSKG 281 - 350 KETIAIRPAAHAEFVPKADLLTLPGAEAGAHGSRRQGKERQGDRSSQSPAGETASISSVSASAEGAVNKV 351 - 420 PLKVIESEKLDEAPEGKRLGFPVHTSVPHTRPETRNKRRAGLPLGGHKSLFLDLPHKVGPDSSQPRGDKK 421 - 490 KPSPPAPAALGKVFNNSASQSSTHKQTSPVPSPLSPRLPSPQQHHRILRLPALPGEREAALNDSPCRKSR 491 - 560 VFSGCVSADTLEPPSSAKVTETKGASPAFLRAGQPRLVPGETLEKSLGPGKTTAEPQHQSPPGISSEGFP 561 - 630 WDGFNEQTPKDLPNRDGGAWVLGYRAGPACPFLLHEEREKSNRSELYLDLHPDHSLTEQDDRTPGRLQAV 631 - 700 WPPPKTKDTEEKVGLKYTEAEYQAAILHLKREHKEEIENLQAQFELRAFHIRGEHAMITARLEETIENLK 701 - 770 HELEHRWRGGCEERKDVCISTDDDCPPKTFRNVCVQTDRETFLKPCESESKTTRSNQLVPKKLNISSLSQ 771 - 840 ...
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(NOTE: Retired. I no longer have the will to continue, and since this is probably the final edit of this thread, I also want to announce that Im leaving...
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Fibrillin microfibrils are polymeric structures within connective cells. believed that fibrillin-1 can compensate generally for the lack of fibrillin-2 which fibrillin-2 can compensate for fibrillin-1 during advancement however, not after P14, when manifestation levels of have become low. Fibrillin-containing microfibrils are polymeric constructions which have been greatest described by their ultrastructural appearance, referred to as the small size nonstriated fibrils connected with flexible materials or with cellar membranes (8) so that as beaded strings, after liberation from cells and rotary shadowing electron microscopy (9). Many structural research of fibrillin-containing microfibrils possess centered on the efforts of fibrillin-1 (10,C14), departing the role of fibrillin-2 undetermined. Nevertheless, some progress continues to be produced using an immunochemical strategy. Antibodies particular for fibrillin-1 and demonstrated that fetal microfibrils are heteropolymeric -2, including both ...
BACKGROUND/AIMS Fascin is an actin-bundling protein that is important in cell motility. Fascin expression has been shown to have a potential role in tumor progression for some epithelial tumors. However, there are only a few studies related to its expression in mesenchymal tumors. We investigated fascin expression in gastrointestinal stromal tumors. METHODS Thirty gastrointestinal stromal tumors, which were very low (n=6), low (n=2), moderate (n=4), and high (n=18) risk, constituted our series. Immunohistochemical expression of fascin was studied in all cases. RESULTS Immunoreactivity was observed in only five cases, all of which were in the high-risk group. The remaining cases (25/30) showed no immunoreactivity, and the difference did not seem statistically important (p=0.261). Fascin expression was stronger in epithelioid cells than spindle-shaped cells (p=0.003). In addition, gastrointestinal stromal tumors in the small bowel showed higher fascin expression than those in the other localizations
The zonular filaments from the eyes of cows are rich in microfibrils containing fibrillin. Tensile tests, stress-relaxation tests and X-ray diffraction studies were used to study the relationship between the mechanical behaviour of zonular filaments and the molecular packing and structure of the fibrillin-rich microfibrils. Zonular filaments show a non-linear (J-shaped) stress-strain curve and appreciable stress-relaxation. It is proposed that the non-linear properties are due to local variations in waviness in the microfibrils or assemblies of microfibrils, which straighten out and become more regularly aligned with strain. Previous and current X-ray diffraction results consistently show a partial ordering of microfibrils in zonular filaments into staggered aggregates which become more ordered and laterally aligned on stretching. Although the removal and re-addition of Ca(2+) is known to change the molecular structure of fibrillin, no effect was observed on the tensile properties of the zonular ...
Allograft inflammatory factor-1 (AIF-1) is an evolutionary conserved protein important to inflammatory responses throughout the body including that of microglia in the central nervous system (CNS). In addition to critical intracellular roles in the activation of microglia and macrophages, AIF-1 can be secreted by these cells in response to inflammatory signals as well as soluble signals released by dying neurons. In response to the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, we found increased levels of AIF-1 expression in cells clustered in the substantia nigra pars compacta (SNpc), the site of dopaminergic cell death. The number of these AIF-1 bright cells continued to increase even after neuronal cell death was complete. This increased expression of AIF-1 was restricted to resident microglia; flow cytometric analysis showed that infiltrating CD45hi leukocytes did not express high levels of AIF-1. Analysis of microglia ex vivo demonstrated the secretion of AIF-1 by these cells, ...
To provide a cellular basis for the increased cutaneous anaphylaxis in Coro1a−/−Coro1b−/− mice, we examined FcεRI-mediated MC degranulation in coronin-deficient BMMCs by measuring the release of β-hexosaminidase from prestored MC granules. Interestingly, antigen-mediated cross-linking of FcεRI resulted in significantly enhanced release of β-hexosaminidase activity from Coro1a−/− BMMCs as compared with WT cells (Fig. 2 b). Although Coro1b−/− BMMCs showed normal degranulation, the additional loss of Coro1b further increased hyperdegranulation in Coro1a−/− BMMCs. For maximal activation, cells were also stimulated with PMA/ionomycin, which overrides the observed hyperdegranulation phenotype of Coro1a−/−Coro1b−/− BMMCs.. Increased FcεRI-mediated degranulation of Coro1a−/− and Coro1a−/−Coro1b−/− BMMCs was further confirmed by assessing the up-regulation of CD107a cell surface expression, which is a marker for granule exocytosis (Fig. 2 c). Again, ...
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