MOLECULAR-CLONING OF HUMAN TESTICULAR ANGIOTENSIN-CONVERTING ENZYME - THE TESTIS ISOZYME IS IDENTICAL TO THE C-TERMINAL HALF OF ENDOTHELIAL ANGIOTENSIN-CONVERTING ENZYME (POLYMERASE CHAIN-REACTION ALTERNATIVE SPLICING)
MOLECULAR-CLONING OF HUMAN TESTICULAR ANGIOTENSIN-CONVERTING ENZYME - THE TESTIS ISOZYME IS IDENTICAL TO THE C-TERMINAL HALF OF ENDOTHELIAL ANGIOTENSIN-CONVERTING ENZYME (POLYMERASE CHAIN-REACTION ALTERNATIVE SPLICING)
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DOI：
10.1073/pnas.86.20.7741
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发表时间：
1989-10-01
影响因子：
11.1
通讯作者：
RIORDAN, JF
中科院分区：
文献类型：
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作者：
EHLERS, MRW;FOX, EA;RIORDAN, JF
血管紧张素转换酶（angiotentin -converting enzyme， ACE; EC 3.4.15.1）是一种广泛存在于哺乳动物组织中的含锌二肽基羧肽酶，被认为在血压调节中起关键作用。睾丸含有一种独特的雄激素依赖性ACE同工酶，功能未知。测定了人睾丸ACE的cDNA序列；它编码一种完全相同的蛋白质，从残基37到C端，再到内皮细胞ACE序列的下半部分或C端结构域[Soubrier， F., Alhenc-Gelas， F., Hubert， C., Allegrini， J., John， M., Tregear， G.和Corvol， P. (1988) Proc. Natl。学会科学。[j]。通过对人睾丸cDNA文库的免疫筛选和杂交筛选，构建了人睾丸ACE全长cDNA。t11和（ii）用ace特异性引物构建并经聚合酶链反应扩增的人睾丸cdna的杂交筛选。推断的蛋白质序列由一个732个残基的前蛋白组成，其中包括一个31个残基的信号肽。成熟多肽的分子量为80,073。睾丸酶含有内皮ACE中两个假定的金属结合位点（His-Glu-Met-Gly-His）中的第二个。这表明功能活性催化位点位于内皮酶的c端结构域内，说明了之前发现这两种结构同工酶的催化作用几乎相同。在克隆和测序的22个睾丸ACE cdna中，有3个具有独特的5”区域，由长达328个碱基对的插入、删除或取代序列组成，这显然是由替代的pre-mRNA剪接引起的。
Angiotensin-converting enzyme (ACE; EC 3.4.15.1) is a zinc-containing dipeptidyl carboxypeptidase widely distributed in mamalian tissues and is thought to play a critical role in blood pressure regulation. Testis contains a unique, androgen-dependent ACE isozyme of unknown function. We have determined the cDNA sequence for human testicular ACE; it encodes a protein that is identical, from residue 37 to its C terminus, to the second half or C-terminal domain of the endothelial ACE sequence [Soubrier, F., Alhenc-Gelas, F., Hubert, C., Allegrini, J., John, M., Tregear, G. and Corvol, P. (1988) Proc. Natl. Acad. Sci. USA 85, 9386-9390]. The full-length human testis ACE cDNA was constructed from a composite of cloned cDNAs, obtained by a combination of (i) immunoscreening and hybridization screening of a human testicular cDNA library in .lambda.t11 and (ii) hybridization screening of human testis cDNAs constructed with ACE-specific primers and amplified by the polymerase chain reaction. The protein sequence inferred consists of a 732-residue preprotein including a 31-residue signal peptide. The mature polypeptide has a molecular weight of 80,073. The testis enzyme contains the second of the two putative metal-binding sites (His-Glu-Met-Gly-His) identified in endothelial ACE. This indicates that the functionally active catalytic site is within the C-terminal domain of the endothelial enzyme, accounting for the previous finding that these two structurally isozymes are virtually identical catalytically. Of 22 testis ACE cDNAs cloned and sequenced, 3 have unique 5'' regions, consisting of inserted, deleted, or substituted sequences up to 328 base pairs long, which have apparently arisen by alternative pre-mRNA splicing.