Hydrolytic reaction by zinc finger mutant peptides: successful redesign of structural zinc sites into catalytic zinc sites.
Hydrolytic reaction by zinc finger mutant peptides: successful redesign of structural zinc sites into catalytic zinc sites.
复制标题
锌指突变肽的水解反应：成功地将结构锌位点重新设计为催化锌位点。
DOI：
10.1021/ic034931y
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发表时间：
2004
影响因子：
4.6
通讯作者：
Y. Sugiura
中科院分区：
文献类型：
--
作者：
A. Nomura;Y. Sugiura
为了将折叠蛋白质结构稳定所需的金属位点重新设计为功能性金属位点，我们构建了一系列锌指突变肽，如zf（CCHG）和zf（GCHH），其中一个锌配位残基被取代为非配位残基。具有与锌离子结合的水的突变肽催化乙酸4-硝基苯酯的水解以及氨基酸酯的对映选择性水解。所有的突变肽的锌络合物显示水解活性，这取决于它们的肽序列。相反，野生型的锌络合物zf（CCHH）和单独的锌离子没有表现出水解能力。这些结果清楚地表明，催化能力主要归因于突变肽的锌络合物中的锌中心。动力学研究表明，催化水解受蛋白质配体的供电子能力和配位环境的影响。此外，水解的pH依赖性强烈表明锌配位的氢氧根离子参与催化反应。这是首次成功研究具有催化活性的锌指肽。
To redesign a metal site originally required for the stabilization of a folded protein structure into a functional metal site, we constructed a series of zinc finger mutant peptides such as zf(CCHG) and zf(GCHH), in which one zinc-coordinating residue is substituted into a noncoordinating one. The mutant peptides having water bound to the zinc ion catalyzed the hydrolysis of 4-nitrophenyl acetate as well as the enantioselective hydrolysis of amino acid esters. All the zinc complexes of the mutant peptides showed hydrolytic activity, depending on their peptide sequences. In contrast, the zinc complex of the wild-type, zf(CCHH), and zinc ion alone exhibited no hydrolytic ability. These results clearly indicate that the catalytic abilities are predominantly attributed to the zinc center in the zinc complexes of the mutant peptides. Kinetic studies of the mutant peptides demonstrated that the catalytic hydrolysis is affected by the electron-donating ability of the protein ligands and the coordination environment. In addition, the pH dependence of the hydrolysis strongly suggests that the zinc-coordinated hydroxide ion participates the catalytic reaction. This report is the first successful study of catalytically active zinc finger peptides.