Ueda M J, Ito T, Ohnishi S, Okada T S
J Cell Sci. 1981 Oct;51:173-88. doi: 10.1242/jcs.51.1.173.
A quantitative method for measuring the initial rate of adhesion of foreign particles to mouse peritoneal macrophages, using protein-coated paraffin oil particles, is described. It was performed to measure the rate of adhesion, which is the first step of phagocytosis, separately from the other phagocytic activities. Kinetic studies showed that the cell-substrate adhesion was a Michaelis-Menten type reaction, which has two rate indices, Km and V. The effects of various reagents on the adhesion rate were analysed. KCN and NaN3 change the rate by affecting V, while the changes in the rate brought about by chlorpromazine and filipin are due to their effect on the Km. Cytochalasin B decreased the adhesion rate, but colchicine did not. The initial rate of adhesion decreased with decreasing temperature. As Arrhenius plot of the initial rate showed a break at 30 degrees C, and gave an activation enthalpy of 14.8 kcal for the process at the physiological temperature. No characteristic change in membrane fluidity was found at 30 degrees C using electron spin resonance spectroscopy.
本文描述了一种使用蛋白包被的石蜡油颗粒来测量外来颗粒与小鼠腹腔巨噬细胞初始黏附速率的定量方法。该方法用于单独测量吞噬作用第一步即黏附速率,以区别于其他吞噬活性。动力学研究表明,细胞 - 底物黏附是米氏(Michaelis - Menten)型反应,有两个速率指标,即Km和V。分析了各种试剂对黏附速率的影响。氰化钾(KCN)和叠氮化钠(NaN3)通过影响V来改变速率,而氯丙嗪和制霉菌素引起的速率变化则是由于它们对Km的影响。细胞松弛素B降低了黏附速率,但秋水仙碱没有。黏附的初始速率随温度降低而下降。初始速率的阿累尼乌斯(Arrhenius)图在30℃处出现转折,在生理温度下该过程的活化焓为14.8千卡。使用电子自旋共振光谱法未发现在30℃时膜流动性有特征性变化。