Abstract
Coated vesicles (CVs), approximately 85 nanomolar in diameter, were obtained from etiolated soybean hypocotyls using discontinuous sucrose gradients. The CVs were treated with 4 molar urea and the vesicle membranes were then removed by centrifugation. When the supernatant subsequently was dialyzed against isolation buffer, sedimentable complexes were obtained. Electron microscopic examination of the pelleted complexes shows many spherical, 65 nanometer baskets consisting of a polygonal lattice free of internal membrane. LDS-PAGE reveals that a 185 kilodalton clathrin heavy chain is enriched in both the basket and CV pellets. However, the overall protein pattern is more complex than that of comparable brain fractions. The results are discussed in terms of the similarities between soybean and brain clathrins and the function of CVs in vivo.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Harrison S. C., Kirchhausen T. Clathrin, cages, and coated vesicles. Cell. 1983 Jul;33(3):650–652. doi: 10.1016/0092-8674(83)90007-7. [DOI] [PubMed] [Google Scholar]
- Joachim S., Robinson D. G. Endocytosis of cationic ferritin by bean leaf protoplasts. Eur J Cell Biol. 1984 May;34(1):212–216. [PubMed] [Google Scholar]
- Kartenbeck J. Preparation of membrane-depleted polygonal coat structures from isolated coated vesicles. Cell Biol Int Rep. 1978 Sep;2(5):457–464. doi: 10.1016/0309-1651(78)90097-8. [DOI] [PubMed] [Google Scholar]
- Keen J. H., Willingham M. C., Pastan I. H. Clathrin-coated vesicles: isolation, dissociation and factor-dependent reassociation of clathrin baskets. Cell. 1979 Feb;16(2):303–312. doi: 10.1016/0092-8674(79)90007-2. [DOI] [PubMed] [Google Scholar]
- Keen J. H., Willingham M. C., Pastan I. Clathrin and coated vesicle proteins Immunological characterization. J Biol Chem. 1981 Mar 10;256(5):2538–2544. [PubMed] [Google Scholar]
- 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]
- Mersey B. G., Fowke L. C., Constabel F., Newcomb E. H. Preparation of a coated vesicle-enriched fraction from plant cells. Exp Cell Res. 1982 Oct;141(2):459–463. doi: 10.1016/0014-4827(82)90235-x. [DOI] [PubMed] [Google Scholar]
- Mueller S. C., Branton D. Identification of coated vesicles in Saccharomyces cerevisiae. J Cell Biol. 1984 Jan;98(1):341–346. doi: 10.1083/jcb.98.1.341. [DOI] [PMC free article] [PubMed] [Google Scholar]
- O'neil R. M., LA Claire J. W., 2nd Mechanical wounding induces the formation of extensive coated membranes in giant algal cells. Science. 1984 Jul 20;225(4659):331–333. doi: 10.1126/science.225.4659.331. [DOI] [PubMed] [Google Scholar]
- Patzer E. J., Schlossman D. M., Rothman J. E. Release of clathrin from coated vesicles dependent upon a nucleoside triphosphate and a cytosol fraction. J Cell Biol. 1982 Apr;93(1):230–236. doi: 10.1083/jcb.93.1.230. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wileman T., Harding C., Stahl P. Receptor-mediated endocytosis. Biochem J. 1985 Nov 15;232(1):1–14. doi: 10.1042/bj2320001. [DOI] [PMC free article] [PubMed] [Google Scholar]