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J Agric Food Chem

Title:		Key Amino Acid Residues Influencing Binding Affinities of Pheromone-Binding Protein from Athetis lepigone to Two Sex Pheromones
Author(s):		Zhang YN; Zhang XQ; Zhang XC; Xu JW; Li LL; Zhu XY; Wang JJ; Wei JY; Mang DZ; Zhang F; Yuan X; Wu XM;
Address:		"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. Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, Tokyo 184-8588, Japan. Key Laboratory of Animal Resistance Research, College of Life Science, Shandong Normal University, Jinan 250100, P. R. China. Institute of Biomedicine, Jinan University, Guangzhou 510000, P. R. China. Zhuhai Trinomab Biotechnology Co., Ltd., Zhuhai 519000, P. R. China"
Journal Title:		J Agric Food Chem
Year:		2020
Volume:		20200519
Issue:		22
Page Number:		6092 - 6103
DOI:		10.1021/acs.jafc.0c01572
ISSN/ISBN:		1520-5118 (Electronic) 0021-8561 (Linking)
Abstract:		"Athetis lepigone 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 E. coli expression system and found it had a higher binding affinity to two sex pheromones of A. lepigone, namely, Z7-12:Ac and Z9-14:Ac (with K(i) 0.77 +/- 0.10 and 1.10 +/- 0.20 muM, 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)"
Keywords:		Amino Acid Motifs Animals Carrier Proteins/chemistry/genetics/*metabolism Female Insect Proteins/chemistry/genetics/*metabolism Kinetics Male Molecular Docking Simulation Moths/chemistry/drug effects/genetics/*metabolism Protein Binding Sex Attractants/ch;
Notes:		"MedlineZhang, Ya-Nan Zhang, Xiao-Qing Zhang, Xiao-Chun Xu, Ji-Wei Li, Lu-Lu Zhu, Xiu-Yun Wang, Juan-Juan Wei, Jun-Yuan Mang, Ding-Ze Zhang, Fan Yuan, Xiaohui Wu, Xiao-Min eng 2020/05/12 J Agric Food Chem. 2020 Jun 3; 68(22):6092-6103. doi: 10.1021/acs.jafc.0c01572. Epub 2020 May 19"