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. 1986 Sep;5(9):2079–2085. doi: 10.1002/j.1460-2075.1986.tb04469.x

Location of the 100 kd-50 kd accessory proteins in clathrin coats.

G P Vigers, R A Crowther, B M Pearse
PMCID: PMC1167085  PMID: 2877872

Abstract

We present a three-dimensional map of the clathrin coat of coated vesicles, generated from tilt series of electron micrographs of unstained specimens embedded in vitreous ice. We have examined native placental coated vesicles and coats reassembled from their purified constituents, namely clathrin triskelions and accessory proteins of approximate mol. wts 100 kd and 50 kd. Our results show that the accessory proteins contribute a further shell of density within the double shell of the clathrin cage, extending from the terminal domains of the clathrin to the membrane of the vesicle. The thickness of the complete coat is approximately 22 nm.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bretscher M. S., Thomson J. N., Pearse B. M. Coated pits act as molecular filters. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4156–4159. doi: 10.1073/pnas.77.7.4156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Crowther R. A., Finch J. T., Pearse B. M. On the structure of coated vesicles. J Mol Biol. 1976 Jun 5;103(4):785–798. doi: 10.1016/0022-2836(76)90209-6. [DOI] [PubMed] [Google Scholar]
  3. Crowther R. A., Pearse B. M. Assembly and packing of clathrin into coats. J Cell Biol. 1981 Dec;91(3 Pt 1):790–797. doi: 10.1083/jcb.91.3.790. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fenner C., Traut R. R., Mason D. T., Wikman-Coffelt J. Quantification of Coomassie Blue stained proteins in polyacrylamide gels based on analyses of eluted dye. Anal Biochem. 1975 Feb;63(2):595–602. doi: 10.1016/0003-2697(75)90386-3. [DOI] [PubMed] [Google Scholar]
  5. Goldstein J. L., Anderson R. G., Brown M. S. Coated pits, coated vesicles, and receptor-mediated endocytosis. Nature. 1979 Jun 21;279(5715):679–685. doi: 10.1038/279679a0. [DOI] [PubMed] [Google Scholar]
  6. Goldstein J. L., Brown M. S., Anderson R. G., Russell D. W., Schneider W. J. Receptor-mediated endocytosis: concepts emerging from the LDL receptor system. Annu Rev Cell Biol. 1985;1:1–39. doi: 10.1146/annurev.cb.01.110185.000245. [DOI] [PubMed] [Google Scholar]
  7. Hanspal M., Luna E., Branton D. The association of clathrin fragments with coated vesicle membranes. J Biol Chem. 1984 Sep 10;259(17):11075–11082. [PubMed] [Google Scholar]
  8. Huang C. Studies on phosphatidylcholine vesicles. Formation and physical characteristics. Biochemistry. 1969 Jan;8(1):344–352. doi: 10.1021/bi00829a048. [DOI] [PubMed] [Google Scholar]
  9. Kirchhausen T., Harrison S. C. Structural domains of clathrin heavy chains. J Cell Biol. 1984 Nov;99(5):1725–1734. doi: 10.1083/jcb.99.5.1725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Lepault J., Pitt T. Projected structure of unstained, frozen-hydrated T-layer of Bacillus brevis. EMBO J. 1984 Jan;3(1):101–105. doi: 10.1002/j.1460-2075.1984.tb01768.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Pearse B. M., Bretscher M. S. Membrane recycling by coated vesicles. Annu Rev Biochem. 1981;50:85–101. doi: 10.1146/annurev.bi.50.070181.000505. [DOI] [PubMed] [Google Scholar]
  13. Pearse B. M. Coated vesicles from human placenta carry ferritin, transferrin, and immunoglobulin G. Proc Natl Acad Sci U S A. 1982 Jan;79(2):451–455. doi: 10.1073/pnas.79.2.451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pearse B. M., Robinson M. S. Purification and properties of 100-kd proteins from coated vesicles and their reconstitution with clathrin. EMBO J. 1984 Sep;3(9):1951–1957. doi: 10.1002/j.1460-2075.1984.tb02075.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Schmid S. L., Matsumoto A. K., Rothman J. E. A domain of clathrin that forms coats. Proc Natl Acad Sci U S A. 1982 Jan;79(1):91–95. doi: 10.1073/pnas.79.1.91. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Stewart M., Vigers G. Electron microscopy of frozen-hydrated biological material. Nature. 1986 Feb 20;319(6055):631–636. doi: 10.1038/319631a0. [DOI] [PubMed] [Google Scholar]
  17. Unanue E. R., Ungewickell E., Branton D. The binding of clathrin triskelions to membranes from coated vesicles. Cell. 1981 Nov;26(3 Pt 1):439–446. doi: 10.1016/0092-8674(81)90213-0. [DOI] [PubMed] [Google Scholar]
  18. Ungewickell E., Branton D. Assembly units of clathrin coats. Nature. 1981 Jan 29;289(5796):420–422. doi: 10.1038/289420a0. [DOI] [PubMed] [Google Scholar]
  19. Ungewickell E., Unanue E. R., Branton D. Functional and structural studies on clathrin triskelions and baskets. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 2):723–731. doi: 10.1101/sqb.1982.046.01.069. [DOI] [PubMed] [Google Scholar]
  20. Vigers G. P., Crowther R. A., Pearse B. M. Three-dimensional structure of clathrin cages in ice. EMBO J. 1986 Mar;5(3):529–534. doi: 10.1002/j.1460-2075.1986.tb04242.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Zaremba S., Keen J. H. Assembly polypeptides from coated vesicles mediate reassembly of unique clathrin coats. J Cell Biol. 1983 Nov;97(5 Pt 1):1339–1347. doi: 10.1083/jcb.97.5.1339. [DOI] [PMC free article] [PubMed] [Google Scholar]

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