Allore R J, Barber B H
Can J Biochem. 1982 Jan;60(1):57-70. doi: 10.1139/o82-009.
The reproducibility of preparation, stability at 4 degrees C, and detergent solubilization characteristics of plasma membrane vesicles purified from domestic pig mesenteric lymph node tissue have been examined. It was found that 2% (w/v) Nonidet P-40 solubilized 50-60% and 2% (w/v) sodium deoxycholate solubilized 60-70% of the total membrane protein. As judged by 125I-labelled lentil lectin staining of the sodium dodecyl sulfate--polyacrylamide gel electrophoresis patterns, 2% (w/v) Nonidet p-40 solubilized approximately 73%, and 2% (w/v) sodium deoxycholate approximately 82% of the total glycoprotein. Actin and a myosin-like component were identified as major constituents of both the Nonidet P-40 and the sodium deoxycholate insoluble fractions, suggesting the possibility that the detergent-insoluble fraction may represent a membrane-associated cytoskeletal network analogous to that which has been demonstrated for the erythrocyte membrane. Consistent with such an intimate association between actin and the plasma membrane, it was found that very little actin could be eluted from the intact membrane vesicles by dialysis against low ionic strength ATP solutions, 0.6 M KCl, or by incubation with DNase I.
对从家猪肠系膜淋巴结组织中纯化的质膜囊泡的制备重现性、4℃下的稳定性以及去污剂增溶特性进行了研究。发现2%（w/v）的Nonidet P-40可增溶50 - 60%的总膜蛋白，2%（w/v）的脱氧胆酸钠可增溶60 - 70%的总膜蛋白。根据十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳图谱上125I标记的扁豆凝集素染色判断，2%（w/v）的Nonidet p-40可增溶约73%的总糖蛋白，2%（w/v）的脱氧胆酸钠可增溶约82%的总糖蛋白。肌动蛋白和一种肌球蛋白样成分被鉴定为Nonidet P-40和脱氧胆酸钠不溶部分的主要成分，这表明去污剂不溶部分可能代表一种与红细胞膜类似的膜相关细胞骨架网络。与肌动蛋白和质膜之间的这种紧密联系一致，发现通过用低离子强度ATP溶液、0.6 M KCl进行透析，或与DNase I孵育，从完整的膜囊泡中几乎洗脱不出肌动蛋白。
