Botti J, Doyennette-Moyne M A, Gouet E, Aubery M, Codogno P
Unité 180 INSERM, Laboratoire de Biologie et Pathologie Moléculaires des Glycoprotéines, Paris, France.
Cell Biol Int Rep. 1987 Dec;11(12):849-59. doi: 10.1016/0309-1651(87)90119-6.
Eight day (8-d CEF) and 16 day old chick embryo fibroblasts (16-d CEF) obtained after a mild trypsin treatment (50 micrograms/ml in Ca2+ and Mg2+-free PBS, plus 10 mM EDTA) for 10 min at 37 degrees C present the same number of fibronectin (FN) binding sites at their surface (approximately 550,000 sites per cell) with a Kd approximately equal to 1.40 microM in both cases. Furthermore, FN interacted with high molecular weight plasma membrane proteins (150,000 and 125,000) insensitive to trypsin treatment. Both 8-d and 16-d CEF adhered and spread to the same extent on a fibronectin coated substratum (80% of the CEF adhered in 60 min). In contrast, 8-d and 16-d CEF behaved differently towards laminin (LM). 8-d CEF exhibited approximately 5500 binding sites per cell with a Kd of 1.5 nM (Codogno P., Doyennette, M.-A. and Aubery M., 1987, Experimental Cell Research, 169, 478-489.) and were highly sensitive to trypsin treatment, whereas 16-d CEF do not express cell surface binding sites for laminin. Differences were also observed in the adhesive capacities of 8-d and 16-d CEF on LM substrata: 8-d CEF adhered and spread on LM in a very specific manner (60% of the cells adhere in 60 min) and 16-d CEF did not adhere to LM even after long periods of incubation exceeding 360 min.
经在37℃下用温和的胰蛋白酶处理(在不含Ca2+和Mg2+的PBS中加入50微克/毫升,再加上10毫摩尔/升的EDTA)10分钟后获得的8日龄(8-d CEF)和16日龄鸡胚成纤维细胞(16-d CEF),其表面的纤连蛋白(FN)结合位点数量相同(每个细胞约550,000个位点),两种情况下解离常数(Kd)均约等于1.40微摩尔。此外,FN与对胰蛋白酶处理不敏感的高分子量质膜蛋白(150,000和125,000)相互作用。8-d和16-d CEF在纤连蛋白包被的基质上的黏附与铺展程度相同(60分钟内80%的CEF黏附)。相比之下,8-d和16-d CEF对层粘连蛋白(LM)的反应不同。8-d CEF每个细胞表现出约5500个结合位点,Kd为1.5纳摩尔(科多尼奥P、多耶内特M.-A.和奥贝里M.,1987年,《实验细胞研究》,169卷,478 - 489页),并且对胰蛋白酶处理高度敏感,而16-d CEF不表达细胞表面的层粘连蛋白结合位点。在8-d和16-d CEF在LM基质上的黏附能力方面也观察到差异:8-d CEF以非常特异的方式在LM上黏附并铺展(60分钟内60%的细胞黏附),而16-d CEF即使在超过360分钟的长时间孵育后也不黏附于LM。