Coated vesicles: characterization, selective dissociation, and reassembly.
AUTOR(ES)
Woodward, M P
RESUMO
Sodium dodecyl sulfate/polyacrylamide gels of coated vesicles from porcine brain (mean 76% coated vesicles) show three major proteins (180,000, 125,000, and 55,000 daltons) that account for 73% of the total protein. Preparations consisting predominantly of coats (65%) have less of the 55,000-dalton protein. Clathrin (180,000 daltons) comprises 40% of the protein of a coated vesicle. Conditions of 2 M urea, 0.25 M MgCl2, or pH 7.5 disrupt the coat and solubilize clathrin. Solubilized clathrin reforms coat structures after dilution of urea or MgCl2. High-pH-solubilized clathrin reassembles after dialysis against buffer at pH 6.5 containing dithiothreitol (5 mM). Reassembled coats are predominantly clathrin.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=336121Documentos Relacionados
- Reassembly of protein-lipid complexes into large bilayer vesicles: Perspectives for membrane reconstitution
- ATP- and Cytosol-dependent Release of Adaptor Proteins from Clathrin-coated Vesicles: A Dual Role for Hsc70
- Dissociation and Reassembly of Soybean Clathrin 1
- Matrix protein in planar membranes: clusters of channels in a native environment and their functional reassembly.
- Yeast Membrane Vesicles: Isolation and General Characteristics1