Analysis of the mechanism of the Serratia nuclease using site-directed mutagenesis
Analysis of the mechanism of the Serratia nuclease using site-directed mutagenesis
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DOI：
10.1093/nar/24.14.2632
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发表时间：
1996-07-15
影响因子：
14.9
通讯作者：
Pingoud, A
中科院分区：
文献类型：
--
作者：
Friedhoff, P;Kolmes, B;Pingoud, A
基于沙雷氏菌核酸酶的晶体结构分析和6个相关核酸酶的序列比对，选择了位于先前鉴定的催化位点残基His89附近的同意氨基酸残基进行诱变研究。在分析的12个氨基酸残基中，有5个对酶的催化活性特别重要：Arg57、Arg87、His89、Asn119和Glu127。例如，它们被丙氨酸取代，导致了活性非常低的突变蛋白，
Based on crystal structure analysis of the Serratia nuclease and a sequence alignment of six related nucleases, consented amino acid residues that are located in proximity to the previously identified catalytic site residue His89 were selected for a mutagenesis study. Five out of 12 amino acid residues analyzed turned out to be of particular importance for the catalytic activity of the enzyme: Arg57, Arg87, His89, Asn119 and Glu127. Their replacement by alanine, for example, resulted in mutant proteins of very low activity,