Robinson G B, Ligler F S
Eur J Biochem. 1980 Oct;111(2):485-90. doi: 10.1111/j.1432-1033.1980.tb04964.x.
Basement membranes were isolated from rabbit kidney using the detergent N-dodecyl sarcosine. Treatment of the membranes with 6 M urea or 5% sodium dodecyl sulphate in the presence of reducing agent at 37 degrees C extracted approximately 50% of the protein. Amino acid analysis and polyacrylamide gel electrophoresis demonstrated that the residual proteins were collagenous and of molecular weight. Extracted membranes retained their continuity of structure and exhibited a matrix composed of fibrous and globular elements when examined by electron microscopy. When the filtration properties of the membranes were studied in vitro they showed an enhanced capacity to retain proteins. The results suggest that basement membranes contain a matrix of fibrous proteins, which is responsible for the sieving characteristics of the native membranes.
使用去污剂N-十二烷基肌氨酸从兔肾中分离出基底膜。在37℃下,在还原剂存在的情况下,用6M尿素或5%十二烷基硫酸钠处理基底膜,可提取约50%的蛋白质。氨基酸分析和聚丙烯酰胺凝胶电泳表明,残留蛋白质具有胶原性且具有分子量。提取后的基底膜保持其结构的连续性,通过电子显微镜检查时,呈现出由纤维状和球状成分组成的基质。当在体外研究基底膜的过滤特性时,它们显示出保留蛋白质的能力增强。结果表明,基底膜含有纤维状蛋白质基质,这是天然膜筛分特性的原因。