人钙调蛋白B (Ca2+/钙调素刺激蛋白磷酸酶的Ca2+结合亚基)cDNA克隆的分离和序列分析。

D Guerini, M H Krinks, J M Sikela, W E Hahn, C B Klee
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引用次数: 48

摘要

我们已经鉴定并克隆了钙调神经磷酸酶的Ca2+结合亚基的人类cDNA,钙调神经磷酸酶是Ca2+/钙调蛋白刺激的蛋白磷酸酶的脑同工酶。2.5 kb的cDNA有一个510 bp的开放阅读框,一个至少500 bp的先导序列和一个1277 bp的3'非编码序列。推导出的人脑钙调磷酸酶B蛋白序列与牛脑钙调磷酸酶B的不同之处在于在羧基末端多了一个缬氨酸,并且Met-11和Ser-153被半胱氨酸取代。还分离出了与氨基酸75 ~ 150相对应的小鼠蛋白的部分克隆。这部分人和小鼠的蛋白序列是相同的,DNA序列具有94%的一致性。人和小鼠各自的mrna大小也相似。从蛋白质水平上观察到,除了HeLa细胞外,脑内mRNA丰度是其他组织的20-60倍,HeLa细胞与脑一样含有丰富的钙调磷酸酶B mRNA。
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Isolation and sequence of a cDNA clone for human calcineurin B, the Ca2+-binding subunit of the Ca2+/calmodulin-stimulated protein phosphatase.

We have identified and cloned human cDNA for the Ca2+-binding subunit of calcineurin, the brain isozyme of the Ca2+/calmodulin-stimulated protein phosphatase. The 2.5-kb cDNA has an open reading frame of 510 bp, a leader sequence of at least 500 bp, and a 1,277-bp 3'-noncoding sequence. The deduced sequence of the human protein differs from bovine brain calcineurin B by an additional valine at the carboxyl terminus and substitution of Met-11 and Ser-153 by cysteine. A partial clone of the mouse protein corresponding to amino acids 75-150 was also isolated. This portion of the human and mouse protein sequence is identical, with the DNA sequences showing 94% identity. The respective mRNAs in human and mouse are also of similar size. As was observed with protein levels, mRNA abundance in brain is 20-60 times that found in other tissues with the exception of HeLa cells which, like brain, contain abundant calcineurin B mRNA.

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