Clathrin‐coated Vesicles: Methods for Preparation

Ernst Ungewickell, Hannover Medical School, Hannover, Germany

Published online: April 2001

DOI: 10.1038/npg.els.0002592

Clathrin-coated vesicles carry proteins and lipids between compartments of eukaryotic cells. Best-studied is their role in receptor-mediated endocytosis and in the transport of lysosomal proteins from the trans-Golgi network to the endosomal system.

Keywords: coated vesicles; clathrin; adaptors; membrane traffic

Figure 1. Analysis of clathrin-coated vesicles by electron microscopy and SDS-PAGE. (a) A negatively stained preparation of clathrin-coated vesicles. Note the characteristic coat that surrounds an internal membrane vesicle of amorphous appearance. (b) The protein composition of highly purified clathrin-coated vesicles from pig brain after SDS-PAGE. (c) A cartoon showing how clathrin triskelia (in red, purple and green) are arranged in the coat surrounding the vesicle. An individual clathrin triskelion with its domains labelled is also shown. The terminal domains (TD) of clathrin often contact transmembrane proteins (R) only indirectly through adaptor proteins as depicted in the inset. Ms C. Rudolph is thanked for contributing this figure.
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 References
    Campbell C, Squicciarini J, Shia M, Pilch P and Fine R (1984) Identification of a protein kinase as an intrinsic component of rat liver coated vesicles. Biochemistry 23: 4420–4426.
    Merisko EM, Farquhar MG and Palade GE (1986) Redistribution of clathrin heavy and light chains in anoxic pancreatic acinar cells. Pancreas 1: 110–123.
    Morris SA, Hannig K and Ungewickell E (1988) Rapid purification of clathrin-coated vesicles by free-flow electrophoresis. European Journal of Cell Biology 47: 251–258.
    Mueller SC and Branton D (1984) Identification of coated vesicles in Saccharomyces cerevisiae. Journal of Cell Biology 98: 341–346.
    Pearse BM (1982) Coated vesicles from human placenta carry ferritin, transferrin, and immunoglobulin G. Proceedings of the National Academy of Sciences of the USA 79: 451–455.
    Rubenstein JL, Fine RE, Luskey BD and Rothman JE (1981) Purification of coated vesicles by agarose gel electrophoresis. Journal of Cell Biology 89: 357–361.
    Woodman PG and Warren G (1991) Isolation of functional, coated, endocytic vesicles. Journal of Cell Biology 112: 1133–1141.
 Further Reading
    Schmid SL (1997) Clathrin-coated vesicle formation and protein sorting: an integrated process. Annual Review of Biochemistry 66: 511–548.

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Cell Compartments and Cellular Organelles | Techniques in Cell Biology
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Article Title: Clathrin‐coated Vesicles: Methods for Preparation
 
How to Cite close
Ungewickell, Ernst(Apr 2001) Clathrin‐coated Vesicles: Methods for Preparation. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0002592]