Gundersen G G, Bulinski J C
Eur J Cell Biol. 1986 Dec;42(2):288-94.
Tyrosinated (Tyr) and detyrosinated (Glu) alpha-tubulins are post-translationally modified species that differ by a single amino acid at their respective C-termini. We have examined the distribution of these two species by immunofluorescence in proliferating and differentiated cells using antisera specifically reactive with each of the forms. In proliferating PtK1 cells, Tyr tubulin was the predominant form in almost every cytoplasmic microtubule (MT); only a few MTs contained detectable Glu tubulin. In contrast, staining of centrioles and primary cilia of PtK1 cells suggested that Glu tubulin was the predominant form in these stable assemblies of MTs. An examination of the distribution (by immunofluorescence) and relative amount (by immunoblot analysis) of the two forms of tubulin in the stable assemblies of MTs present in cultured neuronal cells (neurites), sperm and tracheal cells (axonemes and basal bodies), and platelets and erythrocytes (marginal bands) revealed that, in general, the MTs in these arrays contained substantially elevated levels of Glu tubulin in comparison with the levels in MTs of cultured cells. The one exception, the marginal band of toad erythrocytes, which contained only Tyr tubulin, demonstrates that an elevated level of Glu tubulin is not an obligate feature of a stable array of MTs. Nonetheless, an elevated level of Glu tubulin may be a useful indicator of stable MTs in differentiated cells. It is important to note that commonly used sources of tubulin (e.g., brain or flagella) necessarily yield tubulin that differs strikingly from tubulin of proliferating cells in its content of Glu tubulin.(ABSTRACT TRUNCATED AT 250 WORDS)
酪氨酸化(Tyr)和去酪氨酸化(Glu)的α-微管蛋白是翻译后修饰的产物,它们在各自的C末端仅相差一个氨基酸。我们使用与每种形式特异性反应的抗血清,通过免疫荧光法检测了这两种产物在增殖细胞和分化细胞中的分布。在增殖的PtK1细胞中,Tyr微管蛋白几乎是每个细胞质微管(MT)中的主要形式;只有少数微管含有可检测到的Glu微管蛋白。相反,PtK1细胞中心粒和初级纤毛的染色表明,Glu微管蛋白是这些稳定的微管组装体中的主要形式。对培养的神经元细胞(神经突)、精子和气管细胞(轴丝和基体)以及血小板和红细胞(边缘带)中微管稳定组装体中两种形式微管蛋白的分布(通过免疫荧光)和相对含量(通过免疫印迹分析)进行的检查表明,一般来说,与培养细胞微管中的水平相比,这些阵列中的微管含有显著升高的Glu微管蛋白水平。唯一的例外是蟾蜍红细胞的边缘带,其仅含有Tyr微管蛋白,这表明Glu微管蛋白水平升高并非稳定微管阵列的必然特征。尽管如此,Glu微管蛋白水平升高可能是分化细胞中稳定微管的一个有用指标。需要注意的是,常用的微管蛋白来源(如脑或鞭毛)产生的微管蛋白在其Glu微管蛋白含量上必然与增殖细胞的微管蛋白有显著差异。(摘要截断于250字)