Anhui Province Key Laboratory of Pollutant Sensitive Materials and Environmental Remediation, College of Life Sciences, Huaibei Normal University, Huaibei 235000, P. R. China.
Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, P. R. China.
J Agric Food Chem. 2020 Jun 3;68(22):6092-6103. doi: 10.1021/acs.jafc.0c01572. Epub 2020 May 19.
is a polyphagous pest found around the world that feeds on maize, wheat, and various other important crops. Although it exhibits a degree of resistance to various chemical insecticides, an effective pest-control method has not yet been developed. The sex pheromone communication system plays an essential role in the mating and reproduction of moths, in which pheromone-binding proteins (PBPs) are crucial genes. In this study, we cloned and purified the protein AlepPBP1 using an expression system and found it had a higher binding affinity to two sex pheromones of , namely, Z7-12:Ac and Z9-14:Ac (with 0.77 ± 0.10 and 1.10 ± 0.20 μM, respectively), than to other plant volatiles. The binding-mode analysis of protein conformation with equilibrium stabilization was obtained using molecular dynamics (MD) simulation and indicated that hydrophobic interactions involving several nonpolar residues were the main driving force for the binding affinity of AlepPBP1 with sex pheromones. Computational alanine scanning (CAS) was performed to further identify key amino acid residues and validate their binding contributions. Each key residue, including Phe36, Trp37, Val52, and Phe118, was subsequently mutated into alanine using site-directed mutagenesis. Binding assays showed that the efficient binding abilities to Z7-12:Ac (F36A, W37A, and F118A) and Z9-14:Ac (F36A, W37A, V52A, and F118A) were almost lost in the mutated proteins. Our results demonstrated that these key amino acid residues are crucial for determining the binding ability of AlepPBP1 to sex pheromones. These findings provide a basis for the use of AlepPBP1 in the studies as a specific target for the development of novel behavioral antagonists with marked inhibition or mating-disruption abilities using computer-aided drug design (CADD).
它是一种在世界各地都能找到的多食性害虫，以玉米、小麦和各种其他重要作物为食。尽管它对各种化学杀虫剂表现出一定程度的抗性，但尚未开发出有效的害虫控制方法。性信息素通讯系统在蛾类的交配和繁殖中起着至关重要的作用，其中信息素结合蛋白（PBPs）是关键基因。在这项研究中，我们使用表达系统克隆和纯化了蛋白 AlepPBP1，并发现它对两种 的性信息素有更高的结合亲和力，即 Z7-12:Ac 和 Z9-14:Ac（分别为 0.77±0.10 和 1.10±0.20 μM），而对其他植物挥发物的亲和力较低。通过分子动力学（MD）模拟获得了蛋白构象与平衡稳定的结合模式分析，并表明涉及几个非极性残基的疏水相互作用是 AlepPBP1 与性信息素结合亲和力的主要驱动力。进行了计算丙氨酸扫描（CAS）以进一步确定关键氨基酸残基并验证它们的结合贡献。使用定点突变将每个关键残基，包括 Phe36、Trp37、Val52 和 Phe118，突变为丙氨酸。结合测定表明，突变蛋白对 Z7-12:Ac（F36A、W37A 和 F118A）和 Z9-14:Ac（F36A、W37A、V52A 和 F118A）的有效结合能力几乎丧失。我们的结果表明，这些关键氨基酸残基对于确定 AlepPBP1 与性信息素的结合能力至关重要。这些发现为使用 AlepPBP1 作为特定靶标提供了依据，可用于开发具有显著抑制或交配破坏能力的新型行为拮抗剂，使用计算机辅助药物设计（CADD）。
