Compton D A, Yen T J, Cleveland D W
Department of Biological Chemistry Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
J Cell Biol. 1991 Mar;112(6):1083-97. doi: 10.1083/jcb.112.6.1083.
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 greater than 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.
我们描述了11种单克隆抗体的产生,这些抗体可与人有丝分裂染色体的着丝粒/动粒区域结合。这些抗体是针对有丝分裂染色体支架产生的,并通过对纯化染色体的间接免疫荧光筛选着丝粒/动粒结合情况。用这些抗体进行的免疫印迹分析表明,所有抗原的分子量都大于200 kD,并且是细胞核、染色体和/或染色体支架的组成成分。将这些抗原的免疫定位与着丝粒相关蛋白CENP-B的免疫定位进行比较,发现这些着丝粒/动粒蛋白中的每一种都比CENP-B更位于DNA的外围。在正常经历细胞周期的细胞中,这些抗原表现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。