包被小泡：特性、选择性解离与重新组装

Woodward M P, Roth T F

Proc Natl Acad Sci U S A. 1978 Sep;75(9):4394-8. doi: 10.1073/pnas.75.9.4394.

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.

来自猪脑的包被小泡（平均76%为包被小泡）的十二烷基硫酸钠/聚丙烯酰胺凝胶显示出三种主要蛋白质（180,000、125,000和55,000道尔顿），它们占总蛋白质的73%。主要由包被（65%）组成的制剂中55,000道尔顿的蛋白质较少。网格蛋白（180,000道尔顿）占包被小泡蛋白质的40%。2 M尿素、0.25 M氯化镁或pH 7.5的条件会破坏包被并使网格蛋白溶解。尿素或氯化镁稀释后，溶解的网格蛋白会重新形成包被结构。高pH值溶解的网格蛋白在针对含有二硫苏糖醇（5 mM）的pH 6.5缓冲液进行透析后会重新组装。重新组装的包被主要是网格蛋白。