Clathrin structure characterized with monoclonal antibodies. II. Identification of in vivo forms of clathrin

doi:10.1083/jcb.101.6.2055

This Article

	Full Text (PDF, 1642K)
	Alert me when this article is cited
	Citation Map

Services

	Email this article
	Similar articles in this journal
	Similar articles in PubMed
	Alert me to new content in the JCB
	Download to citation manager

Citing Articles

	Citing Articles via HighWire
	Citing Articles via CrossRef
	Citing Articles via Google Scholar

Google Scholar

	Articles by Brodsky, F. M.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Brodsky, F. M.


Social Bookmarking

	What's this?

ARTICLES

Clathrin structure characterized with monoclonal antibodies. II. Identification of in vivo forms of clathrin

FM Brodsky

Clathrin was isolated from detergent-solubilized, biosynthetically radiolabeled cells by immunoprecipitation with anti-clathrin monoclonal antibodies. Immunoprecipitates obtained after pulse-chase labeling demonstrated that after biosynthesis the LCa light chain of clathrin could be found either complexed to heavy chain or in a free pool (not associated with heavy chain) which decreased steadily over time. More than half of the free LCa disappeared within the first hour after biosynthesis, but some was still detectable after several hours. Incorporation of clathrin LCa light chain and heavy chain into coated vesicles was coordinate and increased up to 4 h after biosynthesis. Comparison of these kinetics suggested that once incorporated into coated vesicles, LCa and heavy chain did not dissociate, even during depolymerization of the vesicle. There was also little apparent degradation of clathrin found in coated vesicles for up to 22 h after biosynthesis. Immunoprecipitation with anti-clathrin monoclonal antibodies was carried out after fractionation of continuously radiolabeled cell lysates using two different sizing columns. These experiments indicated that the triskelion form of clathrin that has been isolated from coated vesicles in vitro also exists in vivo. They also confirmed the existence of a transient but detectable pool of newly synthesized free LCa light chain.
Add to CiteULike CiteULike Add to Complore Complore Add to Connotea Connotea Add to Del.icio.us Del.icio.us Add to Digg Digg Add to Reddit Reddit Add to Technorati Technorati What's this?

This article has been cited by other articles:

Keyel, P. A., Thieman, J. R., Roth, R., Erkan, E., Everett, E. T., Watkins, S. C., Heuser, J. E., Traub, L. M. (2008). The AP-2 Adaptor {beta}2 Appendage Scaffolds Alternate Cargo Endocytosis. Mol. Biol. Cell 19: 5309-5326 [Abstract] [Full Text]
Keyel, P. A., Mishra, S. K., Roth, R., Heuser, J. E., Watkins, S. C., Traub, L. M. (2006). A Single Common Portal for Clathrin-mediated Endocytosis of Distinct Cargo Governed by Cargo-selective Adaptors. Mol. Biol. Cell 17: 4300-4317 [Abstract] [Full Text]
Hinrichsen, L., Meyerholz, A., Groos, S., Ungewickell, E. J. (2006). Bending a membrane: How clathrin affects budding. Proc. Natl. Acad. Sci. USA 103: 8715-8720 [Abstract] [Full Text]
Xiao, J., Dai, R., Negyessy, L., Bergson, C. (2006). Calcyon, a Novel Partner of Clathrin Light Chain, Stimulates Clathrin-mediated Endocytosis. J. Biol. Chem. 281: 15182-15193 [Abstract] [Full Text]
Girard, M., Allaire, P. D., McPherson, P. S., Blondeau, F. (2005). Non-stoichiometric Relationship between Clathrin Heavy and Light Chains Revealed by Quantitative Comparative Proteomics of Clathrin-coated Vesicles from Brain and Liver. Mol. Cell. Proteomics 4: 1145-1154 [Abstract] [Full Text]
Moskowitz, H. S., Yokoyama, C. T., Ryan, T. A. (2005). Highly Cooperative Control of Endocytosis by Clathrin. Mol. Biol. Cell 16: 1769-1776 [Abstract] [Full Text]
Hinrichsen, L., Harborth, J., Andrees, L., Weber, K., Ungewickell, E. J. (2003). Effect of Clathrin Heavy Chain- and {alpha}-Adaptin-specific Small Inhibitory RNAs on Endocytic Accessory Proteins and Receptor Trafficking in HeLa Cells. J. Biol. Chem. 278: 45160-45170 [Abstract] [Full Text]
Jones, A., Spiro, D., Kirchhausen, T, Melancon, P, Wessling-Resnick, M (1999). Studies on the inhibition of endosome fusion by GTPgammaS-bound ARF. J. Cell Sci. 112: 3477-3485 [Abstract]
Lin, H. B., Harley, S. M., Butler, J. M., Beevers, L. (1992). Multiplicity of clathrin light-chain-like polypeptides from developing pea (Pisum sativum L.) cotyledons. J. Cell Sci. 103: 1127-1137 [Abstract]
Brodsky, F. (1988). Living with clathrin: its role in intracellular membrane traffic. Science 242: 1396-1402 [Abstract]
Moore, M., Mahaffey, D., Brodsky, F., Anderson, R. (1987). Assembly of clathrin-coated pits onto purified plasma membranes. Science 236: 558-563 [Abstract]