Wehland J, Sandoval I V
Proc Natl Acad Sci U S A. 1983 Apr;80(7):1938-41. doi: 10.1073/pnas.80.7.1938.
Injection of the alpha, beta-nonhydrolyzable GTP analog, guanosine 5'-[alpha, beta-methylene]triphosphate (pp[CH2]pG) into PtK2, A549, and Swiss 3T3 cells produced dramatic changes in the normal pattern of long radiating microtubules displayed by the cells before injection. Injection of pp[CH2]pG into cells growing in normal medium resulted in the formation of microtubule bundles resistant to depolymerization by Colcemid and calcium. Cells injected with pp[CH2]pG after incubation with Colcemid for 2 hr showed polymerization of tubulin into long wavy ribbons within 2 hr after injection. Removal of Colcemid 1 hr after the injection of pp[CH2]pG resulted in assembly of tubulin into short single randomly oriented microtubules. All cells injected with pp[CH2]pG showed impeded translocation and restriction or absence of intracellular movement. pp[CH2]pG also prevented the fragmentation of Golgi elements in A549 cells treated with Colcemid. Cells first treated with Colcemid and then injected with pp[CH2]pG failed to reassemble the Golgi elements after the removal of Colcemid. Cells in intimate membrane contact with cells injected with pp[CH2]pG showed similar changes in microtubule polymerization, cell movement, and organization of Golgi elements.
向PtK2、A549和瑞士3T3细胞中注射α,β-不可水解的GTP类似物鸟苷5'-[α,β-亚甲基]三磷酸(pp[CH2]pG),会使注射前细胞所呈现的正常长辐射状微管模式发生显著变化。向在正常培养基中生长的细胞注射pp[CH2]pG会导致形成对秋水仙酰胺和钙的解聚具有抗性的微管束。用秋水仙酰胺孵育2小时后再注射pp[CH2]pG的细胞,在注射后2小时内显示微管蛋白聚合成长长的波浪状带。注射pp[CH2]pG 1小时后去除秋水仙酰胺,会导致微管蛋白组装成短的单个随机取向的微管。所有注射了pp[CH2]pG的细胞都显示出转运受阻以及细胞内运动受限或缺乏。pp[CH2]pG还能防止用秋水仙酰胺处理的A549细胞中高尔基体元件的碎片化。先经秋水仙酰胺处理然后注射pp[CH2]pG的细胞,在去除秋水仙酰胺后未能重新组装高尔基体元件。与注射了pp[CH2]pG的细胞紧密膜接触的细胞,在微管聚合、细胞运动和高尔基体元件组织方面表现出类似的变化。