Two single mutations commonly cause qualitative change of nonspecific carboxylesterases in insects
Two single mutations commonly cause qualitative change of nonspecific carboxylesterases in insects
复制标题
两个单一突变通常引起昆虫非特异性羧酸酯酶的质变
DOI：
10.1016/j.ibmb.2010.09.004
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发表时间：
2011-01-01
影响因子：
3.8
通讯作者：
Kang, Le
中科院分区：
文献类型：
--
作者：
Cui, Feng;Lin, Zhe;Kang, Le
羧酸酯酶提供了昆虫对杀虫剂，特别是有机磷(OP)的抗性的关键机制。一种抗性机制是羧酸酯酶性质的质的变化。已观察到两种突变形式G151D和W271L，主要存在于双翅目昆虫中，可影响酶的底物专一性。但这两个单一突变是否会共同改变昆虫羧酸酯酶的性质尚不清楚。本研究克隆了分布于四个目的七种昆虫的羧酸酯酶基因，并对其第151位和271位进行了突变，并在大肠杆菌中表达。纯化的重组蛋白对人工羧酸酯和两种OP杀虫剂的动力学进行了考察。G/A151D和W271L突变分别使87.5%和100%的病例羧酸酯酶活性降低，62.5%和87.5%的病例同时具有OP水解酶活性。因此，271位的改变比151位的改变对底物特异性的影响更有效。这些结果可能表明，这两个突变有可能导致昆虫对杀虫剂产生广泛抗药性。(C)2010爱思唯尔有限公司。保留所有权利。
Carboxylesterases provide key mechanisms of resistance to insecticides, particularly organophosphates (OPs), in insects. One resistance mechanism is a qualitative change in the properties of a carboxylesterase. Two mutant forms, G151D and W271L, have been observed, mostly in dipteran species, to affect substrate specificity of enzymes. But whether these two single mutations can commonly change character of insect carboxylesterases is unknown. In our study carboxylesterase genes from seven insects distributed among four orders were cloned, mutated at position 151 or 271 and expressed in Escherichia coli. The kinetics of the purified recombinant proteins was examined towards an artificial carboxylester and two OP insecticides. The G/A151D and W271L mutation significantly reduced carboxylesterase activity in 87.5% and 100% cases, respectively, and at the same time conferred OP hydrolase activities in 62.5% and 87.5% cases, respectively. Thus, the change at position 271 is more effective to influence substrate specificity than that at position 151. These results may suggest that these two mutations have the potential to cause insecticide resistance broadly in insects. (C) 2010 Elsevier Ltd. All rights reserved.