Perret E, Moudjou M, Geraud M L, Derancourt J, Soyer-Gobillard M O, Bornens M
Département de Biologie Cellulaire et Moléculaire, Laboratoire Arago, CNRS, URA, Banyuls sur mer, France.
J Cell Sci. 1995 Feb;108 ( Pt 2):711-25. doi: 10.1242/jcs.108.2.711.
The monoclonal antibody CTR210 raised against isolated human centrosomes strongly decorates the centrosome and more weakly a domain congruent with the Golgi apparatus in several animal cells (HeLa, 3T3, CHO, PtK2). Both decorations resist Triton extraction in conditions which totally extract the Golgi apparatus, as judged by galactosyltransferase decoration. A 67 kDa centrosomal antigen can be demonstrated in human cells with this antibody. CTR210 also decorates the centrosome or associated structures in several systems, including unicellular eukaryotes such as dinoflagellates or ciliates. A 72 kDa antigen has been identified and purified from the dinoflagellate C. cohnii and its NH2-terminal sequence partially established. It shows a close homology with HSP70 proteins. The possibility that the 72 kDa antigen belongs to this chaperone family was further supported using a mAb reacting, in most species, with HSP70. A polyclonal antibody raised against the 72 kDa antigen from C. cohnii decorates the centrosome in human cells and reacts with the CTR210 centrosomal 67 kDa antigen. These results suggest that specific chaperone proteins are associated with the centrosome in eukaryotic cells. The centrosomal chaperones could participate in the microtubule nucleation reaction or in the process of centrosome assembly.
针对分离出的人中心体产生的单克隆抗体CTR210,能强烈标记几种动物细胞(HeLa细胞、3T3细胞、CHO细胞、PtK2细胞)中的中心体,对与高尔基体一致的区域的标记则较弱。根据半乳糖基转移酶标记判断,在能完全提取高尔基体的条件下,这两种标记都能抵抗Triton提取。用该抗体可在人细胞中检测到一种67 kDa的中心体抗原。CTR210还能标记包括单细胞真核生物如鞭毛虫或纤毛虫在内的多种体系中的中心体或相关结构。已从鞭毛虫C. cohnii中鉴定并纯化出一种72 kDa的抗原,并部分确定了其NH2末端序列。它与HSP70蛋白显示出密切的同源性。使用一种在大多数物种中与HSP70反应的单克隆抗体,进一步支持了72 kDa抗原属于该伴侣蛋白家族的可能性。针对来自C. cohnii的72 kDa抗原产生的多克隆抗体,能标记人细胞中的中心体,并与CTR210中心体67 kDa抗原发生反应。这些结果表明,特定的伴侣蛋白与真核细胞中的中心体相关。中心体伴侣蛋白可能参与微管成核反应或中心体组装过程。