Leivo I, Engvall E
Cancer Research Center, La Jolla Cancer Research Foundation, CA 92037.
Proc Natl Acad Sci U S A. 1988 Mar;85(5):1544-8. doi: 10.1073/pnas.85.5.1544.
We have identified a tissue-specific basement membrane-associated protein by using monoclonal antibodies prepared against a protein fraction of human placenta. In immunofluorescence, the monoclonal antibodies stained basement membranes of Schwann cells, striated muscle, and trophoblast, whereas no reaction was seen with any other basement membrane or tissue structure. In antibody-affinity chromatography of proteolytic digests of human placenta, a 65-kDa polypeptide was bound by these monoclonal antibodies. Rabbit antisera and monoclonal antibodies raised against the isolated 65-kDa polypeptide stained human and monkey tissues identically to the original monoclonal antibodies and reacted with an 80-kDa polypeptide in tissue extracts prepared without proteolysis. The 65-kDa and 80-kDa polypeptides were shown to be immunologically distinct from laminin, type IV collagen, fibronectin, and major serum proteins. They presumably represent a novel basement membrane-associated protein, which we have named merosin. No merosin immunoreactivity could be detected in cultures of any of 28 established cell lines. In developing mouse tissues, merosin staining first appeared at the newborn stage. The restricted tissue distribution and late developmental appearance of merosin suggest that the protein has a tissue-specific function associated with a high level of differentiation.
我们利用针对人胎盘蛋白组分制备的单克隆抗体,鉴定出一种组织特异性的基底膜相关蛋白。在免疫荧光实验中,该单克隆抗体可对施万细胞、横纹肌和滋养层的基底膜进行染色,而对其他任何基底膜或组织结构均无反应。在人胎盘蛋白水解消化产物的抗体亲和层析中,这些单克隆抗体可结合一种65 kDa的多肽。用针对分离出的65 kDa多肽制备的兔抗血清和单克隆抗体,对人和猴组织的染色结果与原始单克隆抗体完全相同,并与未经蛋白水解制备的组织提取物中的一种80 kDa多肽发生反应。结果表明,65 kDa和80 kDa的多肽在免疫上与层粘连蛋白、IV型胶原、纤连蛋白和主要血清蛋白不同。它们可能代表一种新的基底膜相关蛋白,我们将其命名为merosin。在28种已建立的细胞系的培养物中均未检测到merosin免疫反应性。在发育中的小鼠组织中,merosin染色最早出现在新生阶段。merosin有限的组织分布和较晚的发育出现表明,该蛋白具有与高度分化相关的组织特异性功能。