兔Cct6的克隆及Cct复合物在哺乳动物组织中的分布。

G J Schwartz, A M Kittelberger, G B Segel
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引用次数: 4

摘要

Cct6蛋白是参与atp依赖性细胞蛋白折叠的Cct复合体的亚基之一。我们利用人CCT6亚基cDNA从兔肾皮质文库中获得全长cDNA。在兔肾脏和肝脏中分别检测到2和2.5 kb的转录本。兔的CCT6基因与人类基因的一致性为93%;推导出的氨基酸序列有97%相同。对小鼠、兔、人和酵母的Cct6蛋白的系统发育分析表明,Cct6蛋白在物种之间的相似性大于其他Cct亚基。atp结合位点在哺乳动物和酵母中完全保守,支持Cct复合物在atp依赖性蛋白折叠中的作用。利用人Cct6蛋白的多克隆抗体和Western分析,我们发现该亚基在多种兔器官和组织、牛睾丸、人淋巴细胞、人和兔网织细胞以及两种培养的肾细胞系中都有表达。通过Western分析,我们还在兔的几个器官和牛的睾丸中发现了Cct1蛋白。这些数据表征了兔Cct6亚基,并将其与同源物进行了比较。西方的分析支持Cct复合体广泛分布于组织中,在真核生物中高度保守的观点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Cloning of rabbit Cct6 and the distribution of the Cct complex in mammalian tissues.

Cct6 protein is one of the subunits of the Cct complex involved in ATP-dependent folding of cellular proteins. We used the cDNA of the human CCT6 subunit to obtain a full-length cDNA from a rabbit kidney cortex library. Two transcripts of 2 and 2.5 kb were detected in rabbit kidney and liver by Northern analysis. The rabbit CCT6 was 93% identical to the human gene; the deduced amino acid sequence was 97% identical. A phylogenetic analysis of Cct6 proteins from mouse, rabbit, human, and yeast showed greater similarities of Cct6 protein among the species than among other Cct subunits. The ATP-binding sites were perfectly conserved among mammals and yeast, supporting the role of Cct complex in ATP-dependent protein folding. Using a polyclonal antibody to human Cct6 protein and Western analysis, we found expression of this subunit in a variety of rabbit organs and tissues, as well as in bovine testes, human lymphocytes, human and rabbit reticulocytes, and in two cultured kidney cell lines. We also found Cct1 protein by Western analysis in several rabbit organs as well as in bovine testes. These data characterize the rabbit Cct6 subunit and compare it to its homologues. The Western analyses support the concept that Cct complex is widely distributed among tissues and highly conserved among eukaryotes.

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