THE IDENTIFICATION OF MAMMALIAN CENTROSOMAL ANTIGENS USING HUMAN AUTOIMMUNE ANTICENTROSOME ANTISERA
THE IDENTIFICATION OF MAMMALIAN CENTROSOMAL ANTIGENS USING HUMAN AUTOIMMUNE ANTICENTROSOME ANTISERA
复制标题
DOI：
10.1002/cm.970200205
复制
发表时间：
1991-01-01
影响因子：
--
通讯作者：
WEST, K
中科院分区：
文献类型：
--
作者：
BALCZON, R;WEST, K
对人类自身免疫血清进行抗球蛋白自身抗体的筛选。免疫荧光鉴定出两个高滴度的血清与HeLa、CHO和PtK2中心体反应，尽管用这两个抗血清获得的荧光图谱是分开的和不同的。从患者IJ获得的血清含有仅与有丝分裂中心体中存在的表位反应的抗体；使用IJ抗血清从未检测到间期中心体的染色。免疫印迹分析表明，IJ抗血清中的抗体与一个190kD的纺锤体极抗原发生反应。培养的哺乳动物细胞的免疫荧光染色表明，患者SPJ的血清中存在的抗体与间期和有丝分裂的中心体都有反应。SPJ抗血清的免疫印迹鉴定表明，SPJ血清中存在的抗体识别M(R)S 39、185和220kD蛋白，尽管185kD多肽是220kD蛋白的蛋白水解物的可能性尚未被排除。在用SPJ或IJ抗血清预处理的细胞中加入纯6S微管蛋白的裂解细胞系统中，两种抗血清均不能抑制中心体的微管形核。这些抗血清有望成为研究哺乳动物中心体生物化学的有用探针。
Human autoimmune sera were screened for the presence of anticentrosome autoantibodies. Two high titer sera were identified that reacted with HeLa, CHO, and PtK2 centrosomes by immunofluorescence, although the fluorescent patterns that were obtained using the two antisera were separate and distinct. Serum obtained from patient IJ contained antibodies that reacted with epitopes present only in mitotic centrosomes; staining of interphase centrosomes was never detected using IJ antiserum. Immunoblot analysis demonstrated that antibodies present in IJ antiserum reacted with a 190 kD spindle pole antigen. Immunofluorescent staining of cultured mammalian cells demonstrated that antibodies present in serum obtained from patient SPJ reacted with both interphase and mitotic centrosomes. Characterization of SPJ antiserum by immunoblotting demonstrated that antibodies present in the SPJ serum recognized proteins of M(r)S of 39, 185, and 220 kD, although the possibility that the 185 kD polypeptide was a proteolytic breakdown product of the 220 kD protein has not been eliminated. Neither antiserum was able to inhibit microtubule nucleation from centrosomes in a lysed cell system in which pure 6S tubulin was added to permeabilized cells following pretreatment of the cells with either SPJ or IJ antiserum. These antisera should be useful probes for studying the biochemistry of the mammalian centrosome.