Submitted to: Talanta
Publication Type: Peer reviewed journal
Publication Acceptance Date: 1/30/2006
Publication Date: 3/3/2006
Citation: Margaryan, A., Aldrich, J.R., Moaddel, R., Tsuruda, J., Chen, A., Leal, W., Wainer, I. 2006. Synthesis of an immobilized bombyx mori pheromone binding protein liquid chromatography stationary phase.. Talanta. 70(4):752-755. Interpretive Summary: Current approaches to discover new insect repellents are largely based on trial and error. A more logical, and perhaps more fruitful approach, would be to look for chemicals that interfere with the perception of odors that mosquitoes, for example, use to find animals on which to feed. Using the silkworm moth as a model, we found a way to use a cloned version of a small protein found in the antennae of this moth to search for chemicals that affect the moth’s sense of smell. Using this approach, it was possible to precisely study the extent to which other chemical compounds interfered with the perception of the natural pheromone. These experiments show that our method can be used to find new compounds that can jam the sense of smell of insects. This approach is now being tested with other proteins specific to the antennae of female mosquitoes (the only sex that takes a blood meal) in an effort to find new leads for repellents of blood-feeding insects. This information will be of interest to the scientific community studying the process of olfaction and, in particular, researchers seeking new, improved mosquito repellents.
Technical Abstract: The pheromone binding protein from the silkworm moth, Bombyx mori (BmorPBP) has been covalently bonded to a liquid chromatographic stationary phase through either the amino or carboxyl terminal groups. The resulting columns were evaluated using radiolabeled bombykol, and the results demonstrated that the immobilized protein retained its ability to bind this ligand and that the two immobilization techniques produced equivalent columns. The data also demonstrate that the BmorPBP column was able to distinguish between four compounds, and rank them in their relative order of affinity for the protein from highest to lowest: bombykol > bombykal >1-hexadecanol > (Z,E)-5,7-dodecadien-1-ol, and that the immobilized BmorPBP retained its pH-dependent conformational mobility. The column was stable over a 10 month period. The results of this study demonstrate that the concept of immobilizing pheromone binding proteins and odorant binding proteins in order to create affinity chromatographic columns is viable approach to the development of online screens for new insect repellents.