Male tarsi specific odorant-binding proteins in the diving beetle Cybister japonicus sharp

Odorant binding proteins (OBPs) play critical roles in chemical communication of insects, as they recognize and transport environmental chemical signals to receptors. The diving beetle Cybister japonicus Sharp shows a remarkable sexual dimorphism. The foreleg tarsi of males are equipped with large suction cups, believed to help holding the female during underwater courtship and mating. Here, we identified two OBPs highly and specifically expressed in male tarsi, suggesting important functions of these structures in chemical communication. The first protein, Cjap OBP1, exhibits the 6 conserved cysteines motif of classic OBPs, while the second, Cjap OBP2, contains only four cysteines and can be assigned to the sub-class of C-minus OBPs. Both proteins were expressed in a bacterial system and the purified recombinant proteins were used to for antibodies preparation. Western Blot analysis showed that Cjap OBP1 is predominantly expressed in male tarsi and could be also detected in antennae and palpi of both sexes, while Cjap OBP2, besides male tarsi, is also present in testis. Ligand-binding experiments showed a good binding affinity between Cjap OBP1, Cjap OBP2 and citral and coniferyl aldehyde, respectively. These results support a possible function of these two OBPs in the male foreleg tarsi of diving beetles in chemical communication.