ABSTRACT
Odorant binding proteins (OBPs) are required for olfaction perception, and thus may be possible targets for controlling the population of pests by interfering with their chemical communication. A single OBP LmigOBP1 has been identified in the antennae of Locusta migratoria, though four isoforms have been detected. Here, we have investigated the ligand-binding specificity of LmigOBP1 using 67 volatile odor compounds. Fluorescence assays indicate that LmigOBP1 does not bind fecal volatiles or green leaf odors, but shows high affinity for some linear aliphatic compounds, with pentadecanol and 2-pentadecanone being the strongest binding ligands. A 3-dimensional (3D) model of LmigOBP1 was built by homology modeling. Docking simulations based on this model suggested that Asn74 of LmigOBP1 is a key binding site, and this was validated by site-directed mutagenesis and fluorescence assays. We suggest that, as a general rule, a hydrophilic amino acid at the entrance of the binding cavity participates in initial recognition of ligands, and contributes to ligand-binding specificity of OBPs.
Subject(s)
Alcohols/metabolism , Insect Proteins/chemistry , Insect Proteins/metabolism , Locusta migratoria/metabolism , Receptors, Odorant/chemistry , Receptors, Odorant/metabolism , Amino Acid Sequence , Animals , Binding Sites , Insect Proteins/genetics , Kinetics , Ligands , Locusta migratoria/chemistry , Locusta migratoria/genetics , Molecular Sequence Data , Mutagenesis, Site-Directed , Protein Binding , Receptors, Odorant/genetics , Sequence Alignment , Substrate SpecificityABSTRACT
To obtain more information on the elements of chemical communication in the migratory locust (Locusta migratoria) (Orthoptera: Acrididae), we have searched for additional odorant-binding proteins (OBPs) and for volatiles in the feces that could represent potential semiochemicals for this species. A two-dimensional electrophoretic (2DE) analysis of an antennal extract showed only three closely positioned spots that were recognized by the antiserum against locust OBP. Three genes were also identified using PCR and 5'RACE-PCR approaches, encoding isoforms differing from each other for a single amino acid substitution. The gas-chromatographic-electroantennogram (GC-EAD) headspace analysis of a feces sample revealed the presence of several compounds that elicited dose-dependent electrophysiological responses in the antennae of both sexes. Most of these compounds are different from those identified in the feces of the desert locust (Schistocerca gregaria) and reported to be behaviorally active. Ligand-binding experiments performed with such volatiles and recombinant OBP did not show affinity, thus indicating that the binding pocket of OBP requires larger molecules than those so far identified.
