Heterogeneity of microtubule organizing center components as revealed by monoclonal antibodies to mammalian centrosomes and to nucleus-associated bodies from dictyostelium.
Heterogeneity of microtubule organizing center components as revealed by monoclonal antibodies to mammalian centrosomes and to nucleus-associated bodies from dictyostelium.
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哺乳动物中心体和盘基网柄核相关体的单克隆抗体揭示了微管组织中心成分的异质性。
DOI：
10.1002/cm.970220103
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发表时间：
1992
影响因子：
--
通讯作者：
Kuriyama,R
中科院分区：
文献类型：
--
作者：
Sellitto,C;Kimble,M;Kuriyama,R
通过探测针对以下两种单克隆抗体的分子组成，比较了两种形态学上不同的微管组织中心（MTOC）：（i）从细胞黏菌Dictyosteelium discoideum的细胞核复合物中分离的核相关体（NAB）和（ii）从中国仓鼠卵巢（CHO）细胞中分离的哺乳动物有丝分裂纺锤体。用免疫荧光显微镜观察CHO细胞和阿米巴网柄藻的染色模式表明，13种MTOC抗原的分布是不均匀的。不是所有的抗体都能识别间期和有丝分裂中的MTOC。大多数抗MTOC抗体与其他细胞器交叉反应，如细胞核、高尔基体样聚集体和细胞骨架元件。两个抗体，CHO 3和AX3，识别磷酸化的抗原决定簇存在于哺乳动物中心体和DictyosteliumNAB。在免疫印迹上，大多数抗体显示多个条带，通常具有高分子量，表明抗原决定簇在不同分子之间共享。向盖玻片上的去污剂裂解CHO细胞中加入外源性微管蛋白后，一种抗体可抑制微管在体外中心体上的再生长;该抗体可与中心体微管成核活性所必需的抗原结合。这些观察结果表明，在不同的MTOC中的分子组分表现出各种不同的亚细胞定位和功能特性，并且一些抗原分子在形态上不同的MTOC中是保守的。© 1992 Wiley利斯公司
The molecular composition of two morphologically distinct microtubule‐organizing centers (MTOCs) was compared by probing with monoclonal antibodies raised against (i) nucleus‐associated bodies (NABs) isolated in a complex with nuclei from the cellular slime moldDictyostelium discoideumand (ii) mammalian mitotic spindles isolated from Chinese hamster ovary (CHO) cells. The staining patterns observed by immunofluorescence microscopy in whole CHO cells andDictyosteliumamoebae showed that the distribution of thirteen MTOC antigens is heterogeneous. Not all antibodies recognized the MTOC in both interphase and mitosis. Most of the anti‐MTOC antibodies cross‐reacted with other cellular organelles such as nuclei, Golgi apparatus‐like aggregates and cytoskeletal elements. Two antibodies, CHO3 and AX3, recognized phosphorylated epitopes present in both mammalian centrosomes andDictyosteliumNABs. On immunoblots, most of the antibodies showed multiple bands, often of high molecular weight, indicating that the antigenic determinants are shared among different molecules. One antibody inhibited the regrowth of microtubules onto centrosomes in vitro after addition of exogenous tubulin to detergent‐lysed CHO cells on coverslips; this antibody binds to an antigen(s) that might be essential for the microtubule‐nucleating activity of centrosomes. These observations demonstrate that molecular components in different MTOCs exhibit a variety of distinct subcellular localizations and functional properties, and that some antigenic molecules have been conserved among morphologically distinct MTOCs. © 1992 Wiley‐Liss, Inc.