Nabeshima Takeshi, Mori Akio, Kozaki Toshinori, Iwata Yoichi, Hidoh Osamu, Harada Shizuko, Kasai Shinji, Severson David W, Kono Yoshiaki, Tomita Takashi
Institute of Agriculture and Forestry, University of Tsukuba, Tsukuba, Ibaraki 305-8572, Japan.
Biochem Biophys Res Commun. 2004 Jan 16;313(3):794-801. doi: 10.1016/j.bbrc.2003.11.141.
A cDNA sequence encoding a Drosophila Ace-paralogous acetylcholinesterase (AChE) precursor of 701 amino acid residues was identified as the second AChE gene (Ace2) transcript from Culex tritaeniorhynchus. The Ace2 gene is tightly linked to organophosphorus insecticide (OP)-insensitivity of AChE on chromosome 2. The cDNA sequences were compared between an insecticide-susceptible strain and the resistant strain, TYM, that exhibits a 870-fold decrease in fenitroxon-sensitivity of AChE. Two amino acid substitutions were present in TYM mosquitoes. One is F455W whose homologous position in Torped AChE (Phe331) is located in the vicinity of the catalytic His in the acyl pocket of the active site gorge. The other substitution is located to a C-terminal Ile697 position that apparently seems to be excluded from the mature protein and is irrelevant to catalytic activity. The F455W replacement in the Ace2 gene is solely responsible for the insecticide-insensitivity of AChE in TYM mosquitoes.
一个编码701个氨基酸残基的致倦库蚊果蝇Ace同源乙酰胆碱酯酶(AChE)前体的cDNA序列被鉴定为致倦库蚊的第二个AChE基因(Ace2)转录本。Ace2基因与2号染色体上对有机磷杀虫剂(OP)不敏感的AChE紧密连锁。对杀虫剂敏感品系和抗性品系TYM的cDNA序列进行了比较,TYM品系对杀螟硫磷的敏感性降低了870倍。在TYM蚊子中存在两个氨基酸替换。一个是F455W,其在电鳐AChE中的同源位置(Phe331)位于活性位点峡谷酰基口袋中催化性His附近。另一个替换位于C端Ile697位置,该位置显然似乎被排除在成熟蛋白之外,且与催化活性无关。Ace2基因中的F455W替换是导致TYM蚊子中AChE对杀虫剂不敏感的唯一原因。