IDENTIFICATION OF NOVEL CENTROMERE KINETOCHORE-ASSOCIATED PROTEINS USING MONOCLONAL-ANTIBODIES GENERATED AGAINST HUMAN MITOTIC CHROMOSOME SCAFFOLDS
IDENTIFICATION OF NOVEL CENTROMERE KINETOCHORE-ASSOCIATED PROTEINS USING MONOCLONAL-ANTIBODIES GENERATED AGAINST HUMAN MITOTIC CHROMOSOME SCAFFOLDS
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DOI：
10.1083/jcb.112.6.1083
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发表时间：
1991-03-01
影响因子：
7.8
通讯作者：
CLEVELAND, DW
中科院分区：
文献类型：
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作者：
COMPTON, DA;YEN, TJ;CLEVELAND, DW
我们描述了11个单克隆抗体的产生，结合到人类有丝分裂染色体的着丝粒/动粒区域。 这些抗体针对有丝分裂染色体支架产生，并通过针对纯化的染色体的间接免疫荧光筛选着丝粒/动粒结合。 用这些抗体进行的免疫印迹分析显示，所有抗原均> 200 kD，并且是细胞核、染色体和/或染色体支架的组分。 将抗原的免疫定位与观察到的着丝粒相关蛋白CENP-B进行比较，发现这些着丝粒/动粒蛋白中的每一种都比CENP-B更靠近DNA的外周。 在细胞周期正常进展的细胞中，这些抗原显示出四种不同的着丝粒/动粒关联模式，对应于至少四种新的着丝粒/动粒相关蛋白。
We describe the generation of 11 monoclonal antibodies that bind to the centromere/kinetochore region of human mitotic chromosomes. These antibodies were raised against mitotic chromosome scaffolds and screened for centromere/kinetochore binding by indirect immunofluorescence against purified chromosomes. Immunoblot analyses with these antibodies revealed that all of the antigens are > 200 kD and are components of nuclei, chromosomes, and/or chromosome scaffolds. Comparison of the immunolocalization of the antigens with that observed for the centromere-associated protein CENP-B revealed that each of these centromere/kinetochore proteins lies more peripherally to the DNA than does CENP-B. In cells normally progressing through the cell cycle, these antigens displayed four distinct patterns of centromere/kinetochore association, corresponding to a minimum of four novel centromere/kinetochore-associated proteins.