Abstract：
BACKGROUND: General odor-binding proteins (GOBPs) play critical roles in insect olfactory recognition of sex pheromones and plant volatiles. Therefore, the identification of GOBPs in Hyphantria cunea (Drury) based on their characterization to pheromone components and plant volatiles is remain unknown.
RESULTS: In this study, two H. cunea (HcunGOBPs) genes were cloned, and their expression profiles and odorant binding characteristics were systematically analyzed. Firstly, the tissue expression study showed that both HcunGOBP1 and HcunGOBP2 were highly expressed in the antennae of both sexes, indicating their potential involvement in the perception of sex pheromones. Secondly, these two HcunGOBPs genes were expressed in Escherichia coli and ligand binding assays were used to assess the binding affinities to its sex pheromone components including two aldehydes and two epoxides, and some plant volatiles. HcunGOBP2 showed high binding affinities to two aldehyde components (Z9, Z12, Z15-18Ald and Z9, Z12-18Ald), and showed low binding affinities to two epoxide components (1, Z3, Z6-9S, 10R-epoxy-21Hy and Z3, Z6-9S, 10R-epoxy-21Hy), whereas HcunGOBP1 showed weak but significant binding to all four sex pheromone components. Furthermore, both HcunGOBPs demonstrated variable binding affinities to the plant volatiles tested. Thirdly, in silico studies of HcunGOBPs utilized homology, structure modeling, and molecular docking revealed critical hydrophobic residues might be involved in the binding of HcunGOBPs to their sex pheromone components and plant volatiles.
CONCLUSIONS: Our study suggests that these two HcunGOBPs may serve as potential targets for future studies of HcunGOBPs ligand binding, providing insight in the mechanism of olfaction in H. cunea. © 2023 Society of Chemical Industry.
RESULTS: In this study, two H. cunea (HcunGOBPs) genes were cloned, and their expression profiles and odorant binding characteristics were systematically analyzed. Firstly, the tissue expression study showed that both HcunGOBP1 and HcunGOBP2 were highly expressed in the antennae of both sexes, indicating their potential involvement in the perception of sex pheromones. Secondly, these two HcunGOBPs genes were expressed in Escherichia coli and ligand binding assays were used to assess the binding affinities to its sex pheromone components including two aldehydes and two epoxides, and some plant volatiles. HcunGOBP2 showed high binding affinities to two aldehyde components (Z9, Z12, Z15-18Ald and Z9, Z12-18Ald), and showed low binding affinities to two epoxide components (1, Z3, Z6-9S, 10R-epoxy-21Hy and Z3, Z6-9S, 10R-epoxy-21Hy), whereas HcunGOBP1 showed weak but significant binding to all four sex pheromone components. Furthermore, both HcunGOBPs demonstrated variable binding affinities to the plant volatiles tested. Thirdly, in silico studies of HcunGOBPs utilized homology, structure modeling, and molecular docking revealed critical hydrophobic residues might be involved in the binding of HcunGOBPs to their sex pheromone components and plant volatiles.
CONCLUSIONS: Our study suggests that these two HcunGOBPs may serve as potential targets for future studies of HcunGOBPs ligand binding, providing insight in the mechanism of olfaction in H. cunea. © 2023 Society of Chemical Industry.
摘要：
背景：一般气味结合蛋白（GOBP）在昆虫嗅觉识别性信息素和植物挥发物中起关键作用。因此,根据对信息素成分和植物挥发物的表征来鉴定美国白蛾（Drury）中的GOBP仍然未知。
结果：在这项研究中，克隆了两个山葵（HcunGOBPs）基因，并对其表达谱和气味结合特性进行了系统分析。首先,组织表达研究表明，HcunGOBP1和HcunGOBP2在两性的触角中均高表达，表明他们可能参与性信息素的感知。其次,这两个HcunGOBPs基因在大肠杆菌中表达，并使用配体结合测定法来评估与其性信息素成分（包括两种醛和两种环氧化物）的结合亲和力，和一些植物挥发物。HcunGOBP2对两种醛组分（Z9，Z12，Z15-18Ald和Z9，Z12-18Ald）显示出高结合亲和力，并显示出对两种环氧化物成分的低结合亲和力(1，Z3，Z6-9S，10R-环氧-21Hy和Z3，Z6-9S，10R-环氧-21Hy），而HcunGOBP1显示与所有四种性信息素成分的弱但显着结合。此外，两种HcunGOBP均表现出对测试的植物挥发物的可变结合亲和力。第三，在利用同源性的HcunGOBPs的计算机模拟研究中，结构建模,和分子对接显示关键的疏水残基可能参与HcunGOBP与其性信息素成分和植物挥发物的结合。
结论：我们的研究表明，这两种HcunGOBP可能作为未来研究HcunGOBP配体结合的潜在靶标，提供对H.cunea嗅觉机制的见解。本文受版权保护。保留所有权利。
结果：在这项研究中，克隆了两个山葵（HcunGOBPs）基因，并对其表达谱和气味结合特性进行了系统分析。首先,组织表达研究表明，HcunGOBP1和HcunGOBP2在两性的触角中均高表达，表明他们可能参与性信息素的感知。其次,这两个HcunGOBPs基因在大肠杆菌中表达，并使用配体结合测定法来评估与其性信息素成分（包括两种醛和两种环氧化物）的结合亲和力，和一些植物挥发物。HcunGOBP2对两种醛组分（Z9，Z12，Z15-18Ald和Z9，Z12-18Ald）显示出高结合亲和力，并显示出对两种环氧化物成分的低结合亲和力(1，Z3，Z6-9S，10R-环氧-21Hy和Z3，Z6-9S，10R-环氧-21Hy），而HcunGOBP1显示与所有四种性信息素成分的弱但显着结合。此外，两种HcunGOBP均表现出对测试的植物挥发物的可变结合亲和力。第三，在利用同源性的HcunGOBPs的计算机模拟研究中，结构建模,和分子对接显示关键的疏水残基可能参与HcunGOBP与其性信息素成分和植物挥发物的结合。
结论：我们的研究表明，这两种HcunGOBP可能作为未来研究HcunGOBP配体结合的潜在靶标，提供对H.cunea嗅觉机制的见解。本文受版权保护。保留所有权利。
