Bershadsky A D, Gelfand V I
Proc Natl Acad Sci U S A. 1981 Jun;78(6):3610-3. doi: 10.1073/pnas.78.6.3610.
Indirect immunofluorescent staining with an antitubulin antibody was used for studying the role of ATP in the regulation of cytoplasmic microtubule disassembly. Depletion of the cellular ATP pool in cultured mouse fibroblasts with various inhibitors of energy metabolism leads to inhibition of the microtubule disassembly induced by colcemid or vinblastine. Glucose added to the inhibitor-containing incubation medium partially restores the cellular ATP content and abolishes the inhibition of microtubule disassembly. The metabolic inhibitors did not change [3H]colcemid uptake by the cells; therefore, their action on the microtubule disassembly was not caused by the reduction in intracellular colcemid. Addition of ATP to the cytoskeleton preparations obtained by Triton X-100 treatment of the cells markedly stimulates microtubule depolymerization. This effect was specific for ATP; it was not observed in the presence of GTP, UTP, CTP, ADP, AMP, adenosine 5'-(beta, gamma-methylene)triphosphate (a nonhydrolyzable analogue of ATP), or inorganic pyrophosphate or tripolyphosphate. Therefore, depletion of the cellular ATP pool reduces the rate of microtubule disassembly whereas addition of ATP increases it. These results suggest that a certain ATP-dependent reaction [most probably, phosphorylation of some of the microtubule protein(s)] controls microtubule disassembly in the cells.
用抗微管蛋白抗体进行间接免疫荧光染色，以研究ATP在调节细胞质微管解聚中的作用。用各种能量代谢抑制剂使培养的小鼠成纤维细胞中的细胞ATP库耗竭，会导致秋水仙酰胺或长春花碱诱导的微管解聚受到抑制。向含抑制剂的孵育培养基中添加葡萄糖可部分恢复细胞ATP含量，并消除对微管解聚的抑制作用。代谢抑制剂不会改变细胞对[3H]秋水仙酰胺的摄取；因此，它们对微管解聚的作用不是由细胞内秋水仙酰胺的减少引起的。向经Triton X-100处理细胞后获得的细胞骨架制剂中添加ATP，可显著刺激微管解聚。这种效应是ATP特有的；在存在GTP、UTP、CTP、ADP、AMP、腺苷5'-(β,γ-亚甲基)三磷酸（ATP的一种不可水解类似物）、无机焦磷酸或三聚磷酸时未观察到这种效应。因此，细胞ATP库的耗竭会降低微管解聚的速率，而添加ATP则会增加其速率。这些结果表明，某种依赖ATP的反应[很可能是某些微管蛋白的磷酸化]控制着细胞中的微管解聚。
