Pomies P, Block M R
Laboratoire d'Etude des Systèmes Adhésifs Cellulaires, A.T.I.P.E. de l'URA 1178 du CNRS, Université Joseph Fourier, Grenoble, France.
Biochem Biophys Res Commun. 1992 Dec 30;189(3):1429-36. doi: 10.1016/0006-291x(92)90234-c.
In this paper, evidence is provided indicating that the blockade of presynchronized CHO 15B cells in prometaphase by nocodazole is fully reversible and efficient enough to allow us to analyze the function of the integrin receptors. Flow cytometry analysis using a specific antibody raised against the fibronectin receptor, and binding studies of the radiolabeled fibronectin on the cell membrane, indicated a stable number of receptors at the cell surface during mitosis. Furthermore, in the mean time, only a slight increase in the Kd value of the fibronectin-receptor interaction was detected. A binding assay designed to test the affinity of the receptor for its extracellular ligand in an insoluble form was used. No difference was observed between mitotic and interphasic cells. Taken together, these results indicate that the rounding up of the cells observed during mitosis is not due to a loss of the receptor affinity for its extracellular ligand.
本文提供的证据表明,用诺考达唑将预同步化的CHO 15B细胞阻滞在前中期是完全可逆的,并且效率足以让我们分析整合素受体的功能。使用针对纤连蛋白受体产生的特异性抗体进行的流式细胞术分析,以及放射性标记的纤连蛋白在细胞膜上的结合研究表明,有丝分裂期间细胞表面的受体数量稳定。此外,与此同时,仅检测到纤连蛋白-受体相互作用的解离常数(Kd值)略有增加。使用了一种结合试验来测试受体对不溶性细胞外配体的亲和力。在有丝分裂细胞和间期细胞之间未观察到差异。综上所述,这些结果表明,有丝分裂期间观察到的细胞变圆并非由于受体对其细胞外配体的亲和力丧失。