Peptidylprolyl cis/trans isomerase activity and molecular evolution of vertebrate Cyclophilin A
Peptidylprolyl cis/trans isomerase activity and molecular evolution of vertebrate Cyclophilin A
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DOI：
10.16288/j.yczz.15-523
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发表时间：
2016-08-01
期刊：
影响因子：
--
通讯作者：
Sun, Lei
中科院分区：
文献类型：
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作者：
Ren, Liqian;Liu, Wei;Sun, Lei
亲环素A（Cyclophilin A，CypA）是亲环素家族的一个典型成员，参与蛋白质折叠/转位、信号转导、炎症、免疫系统调节、细胞凋亡和病毒复制。本研究对脊椎动物CypA的PPIase活性及其遗传变异进行了研究。根据GenBank参考序列克隆了脊椎动物PPIA基因，其中蝙蝠（Myotis davidi）和鸭（Anas platyrhynchos）的PPM基因为首次报道。将PPIA基因亚克隆到原核表达载体pGEX-6p-1中，并在大肠杆菌中表达。重组CypA蛋白经Sepharose 4 B亲和层析、GST标签切割、凝胶过滤纯化后，PPIase活性测定表明，12种不同脊椎动物CypA蛋白对脯氨酰肽键异构化的催化活性无显著差异。此外，遗传变异和分子进化分析表明，这些脊椎动物CypA蛋白具有相同的CsA结合位点和PPIase活性位点。此外，预测的结构和基因定位是显着保守的。我们的数据表明CypA的重要残基是高度保守的，这是其PPIase活性和细胞功能的关键。
Peptidylprolyl isomerases (PPIase) cyclophilin A (CypA, encoded by PPIA) is a typical member of the Cyclophilin family and is involved in protein folding/translocation, signal transduction, inflammation, immune system regulation, apoptosis and virus replication. In the present study, we investigated the PPIase activity and genetic variation of vertebrate CypA. According to the GenBank reference sequences, vertebrate PPIA genes were cloned, among which the bat (Myotis davidi) and duck (Anas platyrhynchos) PPM genes were reported for the first time. Then PPIA genes were sub-cloned into the expression vector pGEX-6p-1 and expressed in Escherichia coli. Recombinant CypA proteins were purified by using sepharose 4B affinity chromatography and the GST tag was cleaved, followed by gel filtrationi The PPIase activity assay indicated that there was no significant difference in the catalytic activity of prolyl peptide bond isomerization among 12 different vertebrate CypA proteins. In addition, the genetic variation and molecular evolution analysis showed that these vertebrate CypA proteins had the same CsA binding site and the PPIase active sites. Furthermore, the predicted structure and gene localization were remarkable conserved. Our data suggested that the important residues of CypA were highly conserved, which is crucial for its PPIase activity and cellular functions.