Kelleher P J, Juliano R L
J Cell Physiol. 1984 Sep;120(3):329-34. doi: 10.1002/jcp.1041200311.
Chinese hamster ovary (CHO) fibroblasts adhere to the extracellular matrix by both fibronectin-dependent and -independent mechanisms (Harper and Juliano, 1981a,b). Previous studies have suggested that a trypsin-sensitive, 265,000-dalton membrane glycoprotein (gp265) is involved in the fibronectin-independent adhesion process. Using a polyclonal antibody against soluble products obtained from trypsin-treated CHO cells, we have been able to further analyze this involvement. This antibody immunoprecipitates a trypsin-sensitive 265,000-dalton protein from detergent-solubilized cells. Incubation of AdvF11, a variant cell line that does not utilize fibronectin for adhesion, with this antibody blocks their adhesion to extracellular matrix material (ECM). The immunoglobulin fraction will also partially block adhesion of the parental cell line to ECM particularly when the ECM is first treated with an antifibronectin antibody. Taken together these results add support for the involvement of gp265 in fibronectin-independent adhesion and provide a methodology for further characterization.
中国仓鼠卵巢(CHO)成纤维细胞通过纤连蛋白依赖和非依赖机制附着于细胞外基质(Harper和Juliano,1981a,b)。先前的研究表明,一种对胰蛋白酶敏感的265,000道尔顿膜糖蛋白(gp265)参与了非纤连蛋白依赖的黏附过程。使用针对胰蛋白酶处理的CHO细胞获得的可溶性产物的多克隆抗体,我们能够进一步分析这种参与情况。该抗体从去污剂溶解的细胞中免疫沉淀出一种对胰蛋白酶敏感的265,000道尔顿蛋白质。将不利用纤连蛋白进行黏附的变异细胞系AdvF11与该抗体孵育,可阻断它们对细胞外基质材料(ECM)的黏附。免疫球蛋白部分也会部分阻断亲代细胞系对ECM的黏附,特别是当ECM先用抗纤连蛋白抗体处理时。综合这些结果,为gp265参与非纤连蛋白依赖的黏附提供了支持,并为进一步表征提供了一种方法。