title: Effects of mutations on the structure and function of silkworm type 1 acetylcholinesterase.
abstract: AChE is the target of organophosphate (OP) and carbamate (CB) pesticides, and mutations in the gene can significantly reduce insects' sensitivity to these pesticides. Bombyx mori is highly sensitive to pesticides. To investigate the effects of mutations on AChE1 structure and function, we used a prokaryotic system to express B.mori wild type AChE1 (wAChE1) and mutant AChE1 (mAChE1) in this study. Active AChE1 proteins were obtained after refolding and purification, and wAChE1 and mAChE1 had similar activities. After incubation with 10(-6)M physostigmine and 10(-3)mg/mL phoxim, the remaining enzyme activity of mAChE1 was 4.42% and 8.86% higher than that of wAChE1's, respectively. Three-dimensional analysis of mutation AChE1 (mAChE1) revealed that the Ser and Ala side chains extended toward the central part of S285 with distances of just 2.80Å and 3.68Å, respectively, which changed the spatial structure of the active center and reduced its sensitivity to pesticides. These results indicated that the mutations altered the 3D structure of AChE1, which may affect the binding of physostigmine and phoxim to the serine residue at the active center, leading to reduced sensitivity. Our study helps understand the relationship between AChE1 mutations and pesticide resistance and provides a new direction for the cultivation of new pesticide-resistant varieties of B.mori.
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu 215123, PR China.
authors.forename: B B
authors.initials: BB
authors.lastname: Wang
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu 215123, PR China.
authors.forename: F C
authors.initials: FC
authors.lastname: Li
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu 215123, PR China.
authors.forename: K Z
authors.initials: KZ
authors.lastname: Xu
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu 215123, PR China.
authors.forename: M
authors.initials: M
authors.lastname: Ni
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu 215123, PR China.
authors.forename: J S
authors.initials: JS
authors.lastname: Hu
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu 215123, PR China.
authors.forename: J H
authors.initials: JH
authors.lastname: Tian
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu 215123, PR China.
authors.forename: Y Y
authors.initials: YY
authors.lastname: Li
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu 215123, PR China; National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, Jiangsu 215123, PR China.
authors.forename: W D
authors.initials: WD
authors.lastname: Shen
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu 215123, PR China; National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, Jiangsu 215123, PR China. Electronic address: lib@suda.edu.cn.
authors.forename: B
authors.initials: B
authors.lastname: Li
mesh_terms: D000110:Acetylcholinesterase / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D000818:Animals; D012831:Bombyx / Q000201:enzymology*; D008958:Models, Molecular; D009154:Mutation; D010957:Plasmids
chemical_list: D000110:Acetylcholinesterase
keywords: 3D structure; Acetylcholinesterase; Mutations; Phoxim; Physostigmine; Silkworm