Bioinformatic and biochemical analysis of the key binding sites of the pheromone binding protein of Cyrtotrachelus buqueti Guerin-Meneville (Coleoptera: Curculionidea)

• Published in: PeerJ (San Francisco, CA) Vol. 7; p. e7818
• Main Authors: Yang, Hua, Liu, Yan-Lin, Tao, Yuan-Yuan, Yang, Wei, Yang, Chun-Ping, Zhang, Jing, Qian, Li-Zhi, Liu, Hao, Wang, Zhi-Yong
• Format: Journal Article
• Language: English
• Published: San Diego PeerJ. Ltd 14.10.2019; PeerJ, Inc; PeerJ Inc
• Subjects: Analysis; Beetles; Benzene; Binding proteins; Binding sites; Biochemical analysis; Bioinformatics; Bonds (Securities); Camphor; Carbonyl compounds; Chemoreceptors; Cyrtotrachelus buqueti; E coli; Entomology; Fluorescence; Fluorescence assay; Forestry; Hardwoods; Homology; Host plants; Hydrogen; Ligands; Mutants; Pest control; Pheromone binding protein; Pheromones; Plasmids; Protein binding; Proteins; Site-directed mutagenesis; Wang, Chen; Weevils; China
• ISSN: 2167-8359; 2167-8359
• DOI: 10.7717/peerj.7818

The bamboo snout beetle Cyrtotrachelus buqueti is a widely distributed wood-boring pest found in China, and its larvae cause significant economic losses because this beetle targets a wide range of host plants. A potential pest management measure of this beetle involves regulating olfactory chemoreceptors. In the process of olfactory recognition, pheromone-binding proteins (PBPs) play an important role. Homology modeling and molecular docking were conducted in this study for the interaction between CbuqPBP1 and dibutyl phthalate to better understand the relationship between PBP structures and their ligands. Site-directed mutagenesis and binding experiments were combined to identify the binding sites of CbuqPBP1 and to explore its ligand-binding mechanism. The 3D structural model of CbuqPBP1 has six a-helices. Five of these a-helices adopt an antiparallel arrangement to form an internal ligand-binding pocket. When docking dibutyl phthalate within the active site of CbuqPBP1, a CH- π interaction between the benzene ring of dibutyl phthalate and Phe69 was observed, and a weak hydrogen bond formed between the ester carbonyl oxygen and His53. Thus, Phe69 and His53 are predicted to be important residues of CbuqPBP1 involved in ligand recognition. Site-directed mutagenesis and fluorescence assays with a His53Ala CbuqPBP1 mutant showed no affinity toward ligands. Mutation of Phe69 only affected binding of CbuqPBP1 to cedar camphor. Thus, His53 (Between α2 and α3) of CbuqPBP1 appears to be a key binding site residue, and Phe69 (Located at α3) is a very important binding site for particular ligand interactions.