Peters D M, Portz L M, Fullenwider J, Mosher D F
Department of Pathology, University of Wisconsin, Madison 53706.
J Cell Biol. 1990 Jul;111(1):249-56. doi: 10.1083/jcb.111.1.249.
Exogenous plasma and endogenous cellular fibronectins on the surface of cultured fibroblasts and in extracellular matrix fibrils were colocalized by fluorescent and high voltage immunoelectron microscopy. Fibroblast cultures grown in the presence or absence of cycloheximide were incubated with exogenous plasma fibronectin labeled with fluorescein isothiocyanate. A monoclonal antibody specific for the EIIIA sequence of cellular fibronectin was used to detect cellular fibronectin. A rabbit antifluorescein antibody identified fluoresceinated plasma fibronectin. In cultures incubated in the presence of cycloheximide, plasma fibronectin was bound to the cell surface and was assembled into extracellular fibrils. In cultures grown in the absence of cycloheximide, plasma and cellular fibronectins were observed in the same matrix fibrils and in the same locations on the cell surface. There was not, however, random admixture of the two proteins.
通过荧光显微镜和高压免疫电子显微镜观察发现，培养的成纤维细胞表面及细胞外基质纤维中的外源性血浆纤连蛋白和内源性细胞纤连蛋白共定位。在有或无放线菌酮存在的情况下培养成纤维细胞，然后用异硫氰酸荧光素标记的外源性血浆纤连蛋白进行孵育。使用针对细胞纤连蛋白EIIIA序列的单克隆抗体来检测细胞纤连蛋白。用兔抗荧光素抗体鉴定荧光标记的血浆纤连蛋白。在有放线菌酮存在的培养物中，血浆纤连蛋白结合到细胞表面并组装成细胞外纤维。在无放线菌酮培养的细胞中，在相同的基质纤维和细胞表面相同位置观察到血浆纤连蛋白和细胞纤连蛋白。然而，这两种蛋白质并没有随机混合。
