Kreis T E
European Molecular Biology Laboratory, Heidelberg, FRG.
EMBO J. 1987 Sep;6(9):2597-606. doi: 10.1002/j.1460-2075.1987.tb02550.x.
Peptide antibodies specific for tyrosinated (tyr-tubulin) or detyrosinated alpha-tubulin (glu-tubulin) have been generated for studying the relative stability of microtubules enriched in either form of alpha-tubulin. Treatment of Vero cells with nocodazole has revealed that interphase microtubules rich in glu-tubulin (glu-microtubules) are resistant to higher concentrations of the microtubule-disrupting drug than the microtubules containing only tyr-tubulin (tyr-microtubules). Glu-tubulin is enriched in centrioles and mid-bodies, but absent from the first interphase microtubules that have repolymerized in late telophase. Tubulin (including both forms) has been labeled with rhodamine (rh-tubulin) and microinjected into Vero cells to study in vivo the dynamic properties and incorporation rates of tubulin into microtubules rich in either glu- or tyr-tubulin. Tyr-microtubules are significantly more rapidly labeled by the microinjected rh-tubulin than glu-microtubules. Ten minutes after injection, rh-tubulin is present in virtually all tyr-microtubules. The half-time of turnover of glu-microtubules is approximately 1 h. Even several hours after microinjection, some of the glu-microtubules have consistently not incorporated visible amounts of rh-tubulin. These results suggest that tyr- and glu-microtubules respectively represent relatively dynamic and stable subclasses of interphase microtubules.
已制备出针对酪氨酸化(酪氨酸微管蛋白)或去酪氨酸化α-微管蛋白(谷氨酸微管蛋白)的肽抗体,用于研究富含这两种形式α-微管蛋白的微管的相对稳定性。用诺考达唑处理Vero细胞后发现,富含谷氨酸微管蛋白的间期微管(谷氨酸微管)比仅含酪氨酸微管蛋白的微管(酪氨酸微管)对更高浓度的微管破坏药物具有抗性。谷氨酸微管蛋白在中心粒和中间体中富集,但在末期后期重新聚合的首个间期微管中不存在。微管蛋白(包括两种形式)已用罗丹明标记(罗丹明微管蛋白)并显微注射到Vero细胞中,以在体内研究微管蛋白掺入富含谷氨酸或酪氨酸微管蛋白的微管中的动态特性和掺入率。显微注射的罗丹明微管蛋白标记酪氨酸微管的速度明显比标记谷氨酸微管的速度快得多。注射后十分钟,罗丹明微管蛋白几乎存在于所有酪氨酸微管中。谷氨酸微管的周转半衰期约为1小时。即使在显微注射数小时后,一些谷氨酸微管仍始终未掺入可见量的罗丹明微管蛋白。这些结果表明,酪氨酸微管和谷氨酸微管分别代表间期微管中相对动态和稳定的亚类。