Assembly and colchicine binding characteristics of tubulin with maximally tyrosinated and detyrosinated α-tubulins

被引:18
作者
Skoufias, DA [1 ]
Wilson, L [1 ]
机构
[1] Univ Calif Santa Barbara, Dept Mol Cellular & Dev Biol, Santa Barbara, CA 93106 USA
关键词
tubulin tyrosination/detyrosination; colchicine; microtubule dynamics;
D O I
10.1006/abbi.1997.0510
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The posttranslational removal and readdition of tyrosine at the C-terminus of alpha-tubulin is associated with generation of microtubule populations that differ in intracellular distributions, turnover rates, and sensitivities to microtubule-depolymerization agents. Here, we compared the in vitro assembly and colchicine binding characteristics of tubulin dimer preparations composed of alpha-tubulin that had been maximally tyrosinated (similar to 40% tyrosinated) by tubulin-tyrosine ligase and maximally detyrosinated (100% detyrosinated) by carboxypeptidase A. Maximally tyrosinated and detyrosinated tubulins had similar critical concentrations for polymerization and similar association constants for colchicine binding. Microtubules polymerized from the two tubulins also had similar steady-state mean lengths and length distributions. The growing and shortening dynamics (dynamic instability parameters) of individual microtubules made from maximally tyrosinated or detyrosinated Lu-tubulin as determined by video-enhanced dark-field microscopy were similar, but subtle differences in the growing and shortening rates were found. On balance, however, the dynamicity and thus the overall kinetic stability of the two microtubule populations were indistinguishable. The results support the idea that detyrosination of alpha-tubulin does not by itself generate stable microtubules. (C) 1998 Academic Press.
引用
收藏
页码:115 / 122
页数:8
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