一种17-kD着丝粒蛋白（CENP-A）与核小体核心颗粒及组蛋白共同纯化。
A 17-kD centromere protein (CENP-A) copurifies with nucleosome core particles and with histones.
作者信息
Palmer D K, O'Day K, Wener M H, Andrews B S, Margolis R L
出版信息
J Cell Biol. 1987 Apr;104(4):805-15. doi: 10.1083/jcb.104.4.805.
We have detected and begun to characterize a 17-kD centromere-specific protein, CENP-A (Earnshaw, W. C., and N. Rothfield, 1985, Chromosoma., 91:313-321). Sera from several humans with CREST scleroderma autoimmune disease (CREST: calcinosis, Raynaud's phenomenon, esophageal dsymotility, sclerodactyly, and telangiectasia) bind this protein in immunoblot assays of HeLa whole cell or nuclear extracts. We have affinity purified the anti-17-kD centromere protein (anti-CENP-A) specific antibodies from immunoblots of HeLa nuclear protein. The antibodies react with epitopes present on CENP-A derived from humans but apparently do not recognize specific epitopes in either rat or chicken nuclei. Only human nuclear protein is CENP-A positive by immunoblot. Furthermore, human cells show localization of anti-CENP-A antibody to centromeres by immunofluorescence microscopy, whereas rat cells do not. On extraction from the nucleus, CENP-A copurifies with core histones and with nucleosome core particles. We conclude that this centromere-specific protein is a histone-like component of chromatin. The data suggest that CENP-A functions as a centromere-specific core histone.
我们已经检测到并开始对一种17-kD的着丝粒特异性蛋白CENP-A进行特性分析（Earnshaw, W. C., and N. Rothfield, 1985, Chromosoma., 91:313-321）。在对HeLa全细胞或核提取物进行免疫印迹分析时，来自数名患有CREST硬皮病自身免疫疾病（CREST：钙质沉着、雷诺现象、食管运动障碍、指端硬化和毛细血管扩张）的患者血清能与这种蛋白结合。我们已经从HeLa核蛋白的免疫印迹中亲和纯化出了抗17-kD着丝粒蛋白（抗CENP-A）特异性抗体。这些抗体与源自人类的CENP-A上存在的表位发生反应，但显然不能识别大鼠或鸡细胞核中的特异性表位。通过免疫印迹，只有人类核蛋白是CENP-A阳性。此外，通过免疫荧光显微镜观察，人类细胞显示抗CENP-A抗体定位于着丝粒，而大鼠细胞则不然。从细胞核中提取后，CENP-A与核心组蛋白和核小体核心颗粒共纯化。我们得出结论，这种着丝粒特异性蛋白是染色质的一种组蛋白样成分。数据表明CENP-A作为一种着丝粒特异性核心组蛋白发挥作用。
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