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人类桥粒斑珠蛋白(一种常见的连接斑蛋白)的分子克隆及氨基酸序列

Molecular cloning and amino acid sequence of human plakoglobin, the common junctional plaque protein.

作者信息

Franke W W, Goldschmidt M D, Zimbelmann R, Mueller H M, Schiller D L, Cowin P

机构信息

Division of Membrane Biology and Biochemistry, German Cancer Research Center, Heidelberg.

出版信息

Proc Natl Acad Sci U S A. 1989 Jun;86(11):4027-31. doi: 10.1073/pnas.86.11.4027.

Abstract

Plakoglobin is a major cytoplasmic protein that occurs in a soluble and a membrane-associated form and is the only known constituent common to the submembranous plaques of both kinds of adhering junctions, the desmosomes and the intermediate junctions. Using a partial cDNA clone for bovine plakoglobin, we isolated cDNAs encoding human plakoglobin, determined its nucleotide sequence, and deduced the complete amino acid sequence. The polypeptide encoded by the cDNA was synthesized by in vitro transcription and translation and identified by its comigration with authentic plakoglobin in two-dimensional gel electrophoresis. The identity was further confirmed by comparison of the deduced sequence with the directly determined amino acid sequence of two fragments from bovine plakoglobin. Analysis of the plakoglobin sequence showed the protein (744 amino acids; 81,750 Da) to be unrelated to any other known proteins, highly conserved between human and bovine tissues, and characterized by numerous changes between hydrophilic and hydrophobic sections. Only one kind of plakoglobin mRNA (3.4 kilobases) was found in most tissues, but an additional mRNA (3.7 kilobases) was detected in certain human tumor cell lines. This longer mRNA may be represented by a second type of plakoglobin cDNA, which contains an insertion of 297 nucleotides in the 3' non-coding region.

摘要

桥粒斑珠蛋白是一种主要的细胞质蛋白,以可溶性和膜相关形式存在,是已知的两种黏附连接(桥粒和中间连接)的膜下斑块中共有的唯一成分。我们利用牛桥粒斑珠蛋白的部分cDNA克隆,分离出编码人桥粒斑珠蛋白的cDNA,确定其核苷酸序列,并推导其完整的氨基酸序列。通过体外转录和翻译合成了该cDNA编码的多肽,并通过其在双向凝胶电泳中与天然桥粒斑珠蛋白的共迁移进行鉴定。通过将推导序列与牛桥粒斑珠蛋白两个片段的直接测定氨基酸序列进行比较,进一步证实了其同一性。对桥粒斑珠蛋白序列的分析表明,该蛋白(744个氨基酸;81,750道尔顿)与任何其他已知蛋白均无关联,在人和牛组织之间高度保守,其特征是亲水和疏水区域之间有许多变化。在大多数组织中仅发现一种桥粒斑珠蛋白mRNA(3.4千碱基),但在某些人类肿瘤细胞系中检测到另一种mRNA(3.7千碱基)。这种较长的mRNA可能由第二种桥粒斑珠蛋白cDNA代表,该cDNA在3'非编码区含有297个核苷酸的插入。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1798/287381/4ecc5b6fc632/pnas00251-0108-a.jpg

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