Protein engineering of a human enzyme that hydrolyzes V and G nerve agents: design, construction and characterization
Protein engineering of a human enzyme that hydrolyzes V and G nerve agents: design, construction and characterization
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DOI：
10.1016/s0009-2797(99)00053-8
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发表时间：
1999-05-14
影响因子：
5.1
通讯作者：
Millard, CB
中科院分区：
文献类型：
--
作者：
Broomfield, CA;Lockridge, O;Millard, CB
由于目前治疗有机磷抗胆碱酯酶中毒的方法存在不足，我们正试图开发一种可作为预防性保护的催化清除剂。目前已知的酶不能满足这一目的，因为它们结合能力弱，转化率低，所以我们正试图通过蛋白质工程技术来制造合适的酶。丁酰胆碱酯酶突变体G117H具有所需类型的活性，但反应太慢。这种交流描述了一种尝试，以确定反应缓慢的原因，以便可能设计出更有效的酶。结果表明，第117位的突变导致了有机磷化合物与活性部位丝氨酸反应过渡态的扭曲。这些信息将被用来开发避免过渡态稳定位点的其他突变体。爱思唯尔科学爱尔兰有限公司出版。
Because of deficiencies in the present treatments for organophosphorus anticholinesterase poisoning, we are attempting to develop a catalytic scavenger that can be administered as prophylactic protection. Currently known enzymes are inadequate for this purpose because they have weak binding and slow turnover, so we are trying to make an appropriate enzyme by protein engineering techniques. One butyrylcholinesterase mutant, G117H, has the desired type of activity but reacts much too slowly. This communication describes an attempt to determine the reason for the slow reaction so that a more efficient enzyme might be designed. The results indicate that the mutation at residue 117 has resulted in a distortion of the transition state of the reaction of organophosphorus compounds with the active site serine. This information will be used to develop other mutants that avoid transition state stabilization sites. Published by Elsevier Science Ireland Ltd.