Monoclonal antibodies specific for an acetylated form of alpha-tubulin recognize the antigen in cilia and flagella from a variety of organisms.
Monoclonal antibodies specific for an acetylated form of alpha-tubulin recognize the antigen in cilia and flagella from a variety of organisms.
复制标题
DOI：
10.1083/jcb.101.6.2085
复制
发表时间：
1985-12
期刊：
影响因子：
--
通讯作者：
Fuller MT
中科院分区：
文献类型：
--
作者：
Piperno G;Fuller MT
七个单克隆抗体对微管蛋白从海胆精子鞭毛的轴丝识别的乙酰化形式的α-微管蛋白存在于各种生物体的轴丝。在来自多种细胞的可溶性细胞质α-微管蛋白亚型中未检测到抗原。通过粗提物中海胆和衣原体细胞质微管蛋白的体外乙酰化测定抗体的特异性。在提取物中的所有乙酰化多肽中，只有α-微管蛋白具有抗原性。在衣原体微管蛋白同种型中，抗体仅识别在赖氨酸的ε-氨基上体内乙酰化的轴丝α-微管蛋白同种型（L ′ Hernault，S.W.，和J.L. Rosenbaum，1985，Biochemistry，24：473-478）。所述抗体不识别未修饰的轴丝α-微管蛋白、存在于鞭毛基质加膜部分中的未组装的α-微管蛋白或来自衣原体细胞体的可溶性细胞质α-微管蛋白。在含有来自各种来源的轴丝微管的蛋白质组分中发现了抗原，所述来源包括来自海胆囊胚和四膜虫的纤毛、来自果蝇的精子和睾丸以及人类精子。与此相反，抗原中未检测到的可溶性，细胞质微管蛋白，这将不包含微管蛋白从稳定的微管阵列，如中心粒，从未受精的海胆卵，果蝇胚胎，HeLa细胞的制剂。尽管抗体识别的乙酰化α-微管蛋白存在于多种来源的轴丝中，并且可能是轴丝形成所必需的，但它并不专门存在于形态学上不同的轴丝微管的任何一个子集中。抗原被发现在类似的比例从海胆精子轴丝富集中心对或外部双峰B或外部双峰A微管的馏分。因此，α-微管蛋白的乙酰化不能提供任何一类轴丝微管结构的特定机制。初步证据表明，乙酰化α-微管蛋白并不局限于轴丝。本报告中所描述的抗体可能使我们能够推断微管蛋白乙酰化在体内微管结构和功能中的作用。
Seven monoclonal antibodies raised against tubulin from the axonemes of sea urchin sperm flagella recognize an acetylated form of alpha-tubulin present in the axoneme of a variety of organisms. The antigen was not detected among soluble, cytoplasmic alpha-tubulin isoforms from a variety of cells. The specificity of the antibodies was determined by in vitro acetylation of sea urchin and Chlamydomonas cytoplasmic tubulins in crude extracts. Of all the acetylated polypeptides in the extracts, only alpha-tubulin became antigenic. Among Chlamydomonas tubulin isoforms, the antibodies recognize only the axonemal alpha- tubulin isoform acetylated in vivo on the epsilon-amino group of lysine(s) (L'Hernault, S.W., and J.L. Rosenbaum, 1985, Biochemistry, 24:473-478). The antibodies do not recognize unmodified axonemal alpha- tubulin, unassembled alpha-tubulin present in a flagellar matrix-plus- membrane fraction, or soluble, cytoplasmic alpha-tubulin from Chlamydomonas cell bodies. The antigen was found in protein fractions that contained axonemal microtubules from a variety of sources, including cilia from sea urchin blastulae and Tetrahymena, sperm and testis from Drosophila, and human sperm. In contrast, the antigen was not detected in preparations of soluble, cytoplasmic tubulin, which would not have contained tubulin from stable microtubule arrays such as centrioles, from unfertilized sea urchin eggs, Drosophila embryos, and HeLa cells. Although the acetylated alpha-tubulin recognized by the antibodies is present in axonemes from a variety of sources and may be necessary for axoneme formation, it is not found exclusively in any one subset of morphologically distinct axonemal microtubules. The antigen was found in similar proportions in fractions from sea urchin sperm axonemes enriched for central pair or outer doublet B or outer doublet A microtubules. Therefore the acetylation of alpha-tubulin does not provide the mechanism that specifies the structure of any one class of axonemal microtubules. Preliminary evidence indicates that acetylated alpha-tubulin is not restricted to the axoneme. The antibodies described in this report may allow us to deduce the role of tubulin acetylation in the structure and function of microtubules in vivo.