CENP-C, AN AUTOANTIGEN IN SCLERODERMA, IS A COMPONENT OF THE HUMAN INNER KINETOCHORE PLATE
CENP-C, AN AUTOANTIGEN IN SCLERODERMA, IS A COMPONENT OF THE HUMAN INNER KINETOCHORE PLATE
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DOI：
10.1016/0092-8674(92)90538-n
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发表时间：
1992-07-10
期刊：
影响因子：
64.5
通讯作者：
EARNSHAW, WC
中科院分区：
文献类型：
--
作者：
SAITOH, H;TOMKIEL, J;EARNSHAW, WC
我们已经分离并鉴定了一组重叠的互补DNA（cDNA）克隆，它们编码人类着丝粒自身抗原着丝粒蛋白C（CENP - C）。这些克隆的身份是通过几个标准确定的。首先，它们被证明编码一种在十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS - polyacrylamide gel electrophoresis）中迁移至CENP - C预期位置的多肽。其次，我们已经证明这种多肽与人类CENP - C至少有两个表位相同。针对cDNA克隆的非重叠区域编码的融合蛋白制备了多克隆抗体。这些抗体在对人类染色体蛋白进行免疫印迹时，被证明能识别处于CENP - C相应位置的一种蛋白质。此外，我们利用间接免疫荧光法证明这些抗体以符合CENP - C预期模式识别HeLa染色体的着丝粒。通过免疫电子显微镜对CENP - C进行定位显示，这种蛋白是内动粒板的一个组成部分。
We have isolated and characterized a set of overlapping cDNA clones that encode the human centromere autoantigen centromere protein C (CENP-C). The identity of these clones has been established using several criteria. First, they were shown to encode a polypeptide that migrates at the expected position for CENP-C on SDS-polyacrylamide gel electrophoresis. Second, we have demonstrated that this polypeptide shares at least two epitopes with human CENP-C. Polyclonal antibodies were raised to fusion proteins encoded by nonoverlapping regions of the cDNA clones. These antibodies were shown to recognize a protein at a position appropriate for CENP-C on immunoblots of human chromosomal proteins. In addition, we used indirect immunofluorescence to demonstrate that these antibodies recognize centromeres of HeLa chromosomes in the expected pattern for CENP-C. Localization of CENP-C by immunoelectron microscopy reveals that this protein is a component of the inner kinetochore plate.