| Title: | Discrimination of pheromone enantiomers by two pheromone binding proteins from the gypsy moth Lymantria dispar |
| Author(s): | Plettner E; Lazar J; Prestwich EG; Prestwich GD; |
| Address: | "Department of Chemistry, Simon Fraser University, 8888 University Drive, Burnaby, British Columbia V5A 1S6, Canada" |
| ISSN/ISBN: | 0006-2960 (Print) 0006-2960 (Linking) |
| Abstract: | "The gypsy moth, Lymantria dispar, uses (7R, 8S)-cis-2-methyl-7, 8-epoxyoctadecane, (+)-disparlure, as a sex pheromone. The (-) enantiomer of the pheromone is a strong behavioral antagonist. Specialized sensory hairs, sensillae, on the antennae of male moths detect the pheromone. Once the pheromone enters a sensillum, the very abundant pheromone binding protein (PBP) transports the odorant to the sensory neuron. We have expressed the two PBPs found in gypsy moth antennae, PBP1 and PBP2, and we have studied the affinity of these recombinant PBPs for the enantiomers of disparlure. To study pheromone binding under equilibrium conditions, we developed and validated a binding assay. We have addressed the two major problems with hydrophobic ligands in aqueous solution: (1) concentration-dependent adsorption of the ligand on vial surfaces and (2) separation of the protein-bound ligand from the material remaining free in solution. We used this assay to demonstrate for the first time that pheromone binding to PBP is reversible and that the two PBPs from L. dispar differ in their enantiomer binding preference. PBP1 has a higher affinity for the (-) enantiomer, while PBP2 has a higher affinity for the (+) enantiomer. The PBP from the wild silk moth, Antheraea polyphemus (Apol-3) bound the disparlure enantiomers more weakly than either of the L. dispar PBPs, but Apol-3 was also able to discriminate the enantiomers. We have observed extensive aggregation of both L. dispar PBPs and an increase in pheromone binding at high (>2 microM) PBP concentrations. We present a model of disparlure binding to the two PBPs" |
| Keywords: | Alkanes/*metabolism Amino Acid Sequence Animals Carrier Proteins/genetics/*metabolism Insect Proteins/genetics/*metabolism Kinetics Molecular Sequence Data Moths/*metabolism Pheromones/*metabolism Recombinant Proteins/genetics/metabolism Sequence Homology; |
| Notes: | "MedlinePlettner, E Lazar, J Prestwich, E G Prestwich, G D eng Comparative Study Research Support, U.S. Gov't, Non-P.H.S. 2000/07/29 Biochemistry. 2000 Aug 1; 39(30):8953-62. doi: 10.1021/bi000461x" |
