Macrae T H, Ludueña R F
Biochem J. 1984 Apr 1;219(1):137-48. doi: 10.1042/bj2190137.
Tubulin from embryos of the brine shrimp Artemia has been purified to apparent homogeneity by chromatography on phosphocellulose P11 and DEAE-cellulose, (NH4)2SO4 fractionation and assembly-disassembly of microtubules. Peptide mapping indicated that Artemia and bovine brain tubulin were very similar in spite of differences in the electrophoretic behaviour of tubulin from these two organisms. Isoelectric focusing and two-dimensional gel electrophoresis were used to resolve and identify several Artemia isotubulins . The isotubulin composition and the quantity of tubulin did not change during pre-emergence development of Artemia embryos. Formation of microtubules with tubulin purified from embryos at different stages of development did not require glycerol or microtubule-associated proteins and formation of structurally normal microtubules was actually hindered by glycerol and Mg2+. The characteristics of Artemia tubulin, in concert with the unusual life history of Artemia, suggest that this organism will be very useful for the study of tubulin gene expression and tubulin utilization during embryo development.
通过磷酸纤维素P11和DEAE - 纤维素柱层析、硫酸铵分级分离以及微管的组装 - 拆卸,卤虫胚胎中的微管蛋白已被纯化至表观均一。肽图谱分析表明,尽管这两种生物的微管蛋白电泳行为存在差异,但卤虫和牛脑微管蛋白非常相似。等电聚焦和二维凝胶电泳用于分离和鉴定几种卤虫同工微管蛋白。在卤虫胚胎出膜前的发育过程中,同工微管蛋白的组成和微管蛋白的量没有变化。用从不同发育阶段胚胎中纯化的微管蛋白形成微管时,不需要甘油或微管相关蛋白,而甘油和Mg2 +实际上会阻碍结构正常的微管形成。卤虫微管蛋白的特性,与卤虫不同寻常的生活史相一致,表明这种生物对于研究胚胎发育过程中的微管蛋白基因表达和微管蛋白利用将非常有用。