Purification and properties of 100-kd proteins from coated vesicles and their reconstitution with clathrin - PubMed (original) (raw)

Purification and properties of 100-kd proteins from coated vesicles and their reconstitution with clathrin

B M Pearse et al. EMBO J. 1984 Sep.

Abstract

Bullock brain coated vesicles contain a family of at least six 100-kd polypeptides which have the property of promoting clathrin assembly. These proteins have been purified from Triton X-100-extracted coated vesicles by a combination of gel filtration and chromatography on hydroxylapatite and DE-52 cellulose. Three major 100-kd species occur as complexes with a stoichiometric amount of a 50-kd polypeptide. On cross-linking these complexes, the chief products appear to contain two polypeptides of 100 kd and two of 50 kd. These 100-kd/50-kd complexes will polymerise with low concentrations of clathrin to give a relatively homogeneous population of coats predominantly of the 'barrel' size. In contrast, three other polypeptides of 100 kd lack the 50-kd protein but polymerise with clathrin under the same conditions to yield coats of a wide range of sizes including 'barrels', truncated icosahedra and particles of greater than 100 nm diameter. When clathrin cages are reassembled with a saturating amount of 100-kd/50-kd complexes and studied by electron microscopy, the additional proteins appear to follow the underlying geometry of the clathrin polyhedra, partially filling in the polygonal faces of the cage structures. Saturation appears to require approximately 3 molecules of 100-kd polypeptide per clathrin trimer.

PubMed Disclaimer

Cited by

Conserved and plant-unique mechanisms regulating plant post-Golgi traffic.
Fujimoto M, Ueda T. Fujimoto M, et al. Front Plant Sci. 2012 Aug 28;3:197. doi: 10.3389/fpls.2012.00197. eCollection 2012. Front Plant Sci. 2012. PMID: 22973281 Free PMC article.
Auxilin, a newly identified clathrin-associated protein in coated vesicles from bovine brain.
Ahle S, Ungewickell E. Ahle S, et al. J Cell Biol. 1990 Jul;111(1):19-29. doi: 10.1083/jcb.111.1.19. J Cell Biol. 1990. PMID: 1973169 Free PMC article.
Structural and functional division into two domains of the large (100- to 115-kDa) chains of the clathrin-associated protein complex AP-2.
Kirchhausen T, Nathanson KL, Matsui W, Vaisberg A, Chow EP, Burne C, Keen JH, Davis AE. Kirchhausen T, et al. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2612-6. doi: 10.1073/pnas.86.8.2612. Proc Natl Acad Sci U S A. 1989. PMID: 2495531 Free PMC article.
Evolving nature of the AP2 alpha-appendage hub during clathrin-coated vesicle endocytosis.
Praefcke GJ, Ford MG, Schmid EM, Olesen LE, Gallop JL, Peak-Chew SY, Vallis Y, Babu MM, Mills IG, McMahon HT. Praefcke GJ, et al. EMBO J. 2004 Nov 10;23(22):4371-83. doi: 10.1038/sj.emboj.7600445. Epub 2004 Oct 21. EMBO J. 2004. PMID: 15496985 Free PMC article.
A large-scale conformational change couples membrane recruitment to cargo binding in the AP2 clathrin adaptor complex.
Jackson LP, Kelly BT, McCoy AJ, Gaffry T, James LC, Collins BM, Höning S, Evans PR, Owen DJ. Jackson LP, et al. Cell. 2010 Jun 25;141(7):1220-9. doi: 10.1016/j.cell.2010.05.006. Cell. 2010. PMID: 20603002 Free PMC article.

References

1. EMBO J. 1983;2(8):1393-400 - PubMed
1. EMBO J. 1983;2(8):1401-8 - PubMed
1. Nature. 1970 Aug 15;227(5259):680-5 - PubMed
1. Biochem J. 1973 Dec;135(4):867-73 - PubMed
1. Anal Biochem. 1975 Feb;63(2):595-602 - PubMed

Purification and properties of 100-kd proteins from coated vesicles and their reconstitution with clathrin - PubMed (original) (raw)