Ohno M, Martinez-Hernandez A, Ohno N, Kefalides N A
Biochem Biophys Res Commun. 1983 May 16;112(3):1091-8. doi: 10.1016/0006-291x(83)91730-8.
Laminin components were solubilized from basement membranes of amnion, chorion and chorionic microvessels of human placenta without prior protease digestion. These structures, after isolation, were initially processed in a sonicator bath containing a solution of Triton X-100, EDTA and 2M NaCl and the laminins extracted sequentially with 0.5M NaCl, 8 M urea, and 8 M urea + 2% 2-mercaptoethanol + 2% SDS. A high molecular weight (appr. 1 x 10(6)) complex containing laminins was purified by gel filtration on a Sepharose CL-2B column. This complex migrated as a single band on gel electrophoresis before reduction but resolved, after reduction, into four major laminin components, laminin A (350,000 M.W.), laminin M (240,000 M.W.) and laminins B1 and B2 (195,000 and 185,000 M.W.). Laminin M is a new molecular species of this protein.
层粘连蛋白成分从人胎盘羊膜、绒毛膜和绒毛膜微血管的基底膜中溶解出来，无需事先进行蛋白酶消化。这些结构分离后，首先在含有Triton X-100、EDTA和2M NaCl溶液的超声浴中处理，然后依次用0.5M NaCl、8M尿素以及8M尿素+2% 2-巯基乙醇+2% SDS提取层粘连蛋白。通过在Sepharose CL-2B柱上进行凝胶过滤，纯化出一种含有层粘连蛋白的高分子量（约1×10⁶）复合物。该复合物在还原前在凝胶电泳上迁移为单一条带，但还原后分解为四种主要的层粘连蛋白成分，即层粘连蛋白A（分子量350,000）、层粘连蛋白M（分子量240,000）以及层粘连蛋白B1和B2（分子量分别为195,000和185,000）。层粘连蛋白M是该蛋白的一种新分子类型。
