Tomita M, Ishikawa H
Department of Biochemistry, Saitama Medical School, Japan.
Biol Cell. 1992;76(1):103-9. doi: 10.1016/0248-4900(92)90200-k.
When mouse peritoneal macrophages were made to adhere firmly on glass surface and then removed by sequential treatment with hypotonic triethanolamine and Nonidet P-40, a set of proteins were found to be left behind at the sites of adherent cells. Such glass-adherent proteins were detected as round or ellipsoidal patches of autofluorescence under a confocal laser microscope, and visualized ultrastructurally as aggregates of narrow threads of unique loop structures which were composed of linearly aligned particles of 22 +/- 2 nm in diameter. Lithium dodecylsulfate-polyacrylamide gel electrophoresis of the glass-adherent proteins showed two major bands, 12 kDa and 14 kDa, which always co-existed in any different sample. The polyclonal antibody raised against these two proteins specifically stained the glass-adherent proteins in situ. The adhesion of macrophages to glass was significantly blocked with Fab fragments of the antibody. The in situ cross-linking experiment suggested that these two proteins might be closely associated with each other to form complexes. Hence, these proteins can be reasonably considered to be responsible for non-specific adhesion of macrophages to glass.
当使小鼠腹腔巨噬细胞牢固地粘附在玻璃表面，然后通过用低渗三乙醇胺和诺乃洗涤剂P-40顺序处理将其去除时，发现一组蛋白质留在粘附细胞的部位。在共聚焦激光显微镜下，此类玻璃粘附蛋白被检测为自发荧光的圆形或椭圆形斑块，超微结构上显示为独特环状结构的窄线聚集体，这些窄线由直径为22±2nm的线性排列颗粒组成。玻璃粘附蛋白的十二烷基硫酸锂-聚丙烯酰胺凝胶电泳显示出两条主要条带，12kDa和14kDa，它们在任何不同样品中总是同时存在。针对这两种蛋白质产生的多克隆抗体可特异性地原位染色玻璃粘附蛋白。抗体的Fab片段可显著阻断巨噬细胞与玻璃的粘附。原位交联实验表明这两种蛋白质可能彼此紧密关联形成复合物。因此，这些蛋白质可合理地被认为负责巨噬细胞与玻璃的非特异性粘附。
