The non-target toxicity and resistance problems of acetylcholinesterase (AChE) insecticides, such as organophosphates and carbamates, are of growing concern. To explore the potential targets for achieving inhibitor selectivity, the AChE structures at or near the catalytic pocket of Tetranychus urticae (TuAChE), honey bees, and humans were compared. The entrances to the AChE catalytic pocket differ significantly because of their different peripheral sites. The role of these potential mite-specific sites in AChE function was further elucidated by site-directed mutagenesis of these sites and then examining the catalytic activities of TuAChE mutants. The spider mite E₃₁₆, H₃₆₉, and V₁₀₅ active sites are important for AChE function. By further analyzing their physostigmine inhibitory properties and the detailed interaction between physostigmine and TuAChE, the peripheral site H₃₆₉ locating near the gorge entrance, and S₁₅₄ at the oxyanion hole, affects substrate and inhibitor trafficking. The discovery of conserved mite-specific residues in Tetranychus will enable the development of safer, effective pesticides that target residues present only in mite AChEs, potentially offering effective control against this important agricultural pest.

乙酰胆碱酯酶（AChE）杀虫剂（如有机磷酸酯和氨基甲酸酯）的非目标毒性和耐药性问题日益引起人们的关注。为了探索实现抑制剂选择性的潜在目标，比较了四叶草（Tutranychus urticae，TuAChE），蜜蜂和人类的催化口袋处或附近的AChE结构。AChE催化袋的入口因周边位置不同而有很大差异。这些潜在的螨特异位点在AChE功能中的作用通过这些位点的定向诱变进一步阐明，然后研究了TuAChE突变体的催化活性。红蜘蛛E ₃₁₆，H ₃₆₉和V ₁₀₅活动位点对于AChE功能很重要。通过进一步分析它们的毒扁豆碱抑制特性和毒扁豆碱与TuAChE之间的详细相互作用，外围位点H ₃₆₉位于峡谷入口附近，S ₁₅₄位于氧阴离子孔处，影响底物和抑制剂的运输。在Tetranychus中发现保守的螨特有残留物将使开发更安全，有效的农药成为目标，这些农药仅针对仅存在于螨类AChE中的残留物，从而有可能对这种重要的农业害虫提供有效的控制。