Coated vesicles: characterization, selective dissociation, and reassembly.
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.
D000818:Animals; D001921:Brain / Q000648:ultrastructure; D002458:Cell Fractionation / Q000379:methods; D003594:Cytoplasmic Granules / Q000032:analysis / Q000648:ultrastructure*; D006863:Hydrogen-Ion Concentration; D007425:Intracellular Membranes / Q000032:analysis / Q000648:ultrastructure; D008274:Magnesium / Q000494:pharmacology; D008565:Membrane Proteins / Q000032:analysis*; D008970:Molecular Weight; D013552:Swine; D014508:Urea / Q000494:pharmacology
D008565:Membrane Proteins; D014508:Urea; D008274:Magnesium