title: Ligands binding and molecular simulation: the potential investigation of a biosensor based on an insect odorant binding protein.
abstract: Based on mimicking biological olfaction, biosensors have been applied for the detection of various ligands in complex environment, which could represent one of the most promising research fields. In this study, the basic characters of one insect odorant binding protein (OBP) as a biosensor were explored. To explore the molecular recognition process, the tertiary structure of the protein was modeled and the protein-ligand interactions with 1,536,550 chemicals were investigated by the molecular docking. The availability of large amount of recombinant SlitOBP1 overcame the difficulty to obtain biological sensing material. After obtained the purified recombinant protein, the result of fluorescence binding assays proved the candidate protein has good affinities with the majority of the tested chemicals. With the aid of simulation docking, the key conserved amino acids within the binding site were identified and then mutated to alanine. After mutation, the protein-ligand binding characteristics were recorded, and the competitive binding assays were carried out to provide experimental verification. The detailed information on its structure and affinities investigated in this study could allow the design of specific mutants with desired characteristics, which provides a solid base for tailoring OBP for biosensor and provides a role model for screening the other elements in olfactory system for different applications.
authors.affiliation: Laboratory of Insect Toxicology, Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou, People's Republic of China.
authors.forename: Xin
authors.initials: X
authors.lastname: Yi
authors.affiliation: Laboratory of Insect Toxicology, Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou, People's Republic of China.
authors.forename: Yanbo
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Laboratory of Insect Toxicology, Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou, People's Republic of China.
authors.forename: Peidan
authors.initials: P
authors.lastname: Wang
authors.affiliation: Laboratory of Insect Toxicology, Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou, People's Republic of China.
authors.forename: Jiangwei
authors.initials: J
authors.lastname: Qi
authors.affiliation: Laboratory of Insect Toxicology, Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou, People's Republic of China.
authors.forename: Meiying
authors.initials: M
authors.lastname: Hu
authors.affiliation: Laboratory of Insect Toxicology, Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou, People's Republic of China.
authors.forename: Guohua
authors.initials: G
authors.lastname: Zhong
mesh_terms: D000818:Animals; D001665:Binding Sites / Q000235:genetics; D032701:Biomimetics / Q000379:methods*; D015374:Biosensing Techniques / Q000379:methods*; D019295:Computational Biology; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D005453:Fluorescence; D008024:Ligands; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D011487:Protein Conformation; D018035:Receptors, Odorant / Q000737:chemistry* / Q000378:metabolism*; D018411:Spodoptera / Q000235:genetics / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D008024:Ligands; D018035:Receptors, Odorant; C045648:odorant-binding protein
keywords: Biosensors; Fluorescence binding; Molecular simulation; Odorant binding protein; Site-directed mutagenesis.