Spangenberg P, Heptinstall S, Crawford J, Till U
Institute of Pathological Biochemistry, Medical Academy of Erfurt, Germany.
Acta Histochem Suppl. 1991;41:181-6.
The amounts of the different forms of actin (G-actin, F-actin) can be measured biochemically following lysis of the cells by the DNase I inhibition assay. The existing methodology for studying G-actin in unstimulated platelets was found to be inappropriate for studies during ADP-induced platelet activation. However, this problem was overcome by a simple modification of the procedure in which formaldehyde was added to the buffer used to lyse the activated platelets. Using this modification the G-actin values obtained immediately after lysis did not change during storage of the lysates on ice for more than 30 minutes. The results show rapid conversion of G-actin to F-actin in association with the shape change during ADP-stimulated activation. Examination of unstimulated platelets using the modified procedure enabled identification of a pool of actin that is rapidly dissociated to G-actin in the absence of formaldehyde, the existence of which had not previously been recognised.
通过DNase I抑制测定法裂解细胞后，可对不同形式的肌动蛋白（G-肌动蛋白、F-肌动蛋白）的含量进行生化测定。现有的用于研究未受刺激血小板中G-肌动蛋白的方法被发现不适用于ADP诱导的血小板活化过程中的研究。然而，通过对该程序进行简单修改，即在用于裂解活化血小板的缓冲液中添加甲醛，这个问题得到了解决。使用这种修改方法，裂解后立即获得的G-肌动蛋白值在裂解物于冰上储存超过30分钟的过程中不会改变。结果表明，在ADP刺激的活化过程中，伴随着形状变化，G-肌动蛋白迅速转化为F-肌动蛋白。使用修改后的程序对未受刺激的血小板进行检测，能够识别出在没有甲醛的情况下会迅速解离为G-肌动蛋白的肌动蛋白池，其存在此前未被认识到。
