cDNA cloning and characterization of the warm-temperature-acclimation-associated protein Wap65 from carp, Cyprinus carpio

We determined full-length cDNA of carp warm-temperature-acclimation-associa ted 65-kDa protein (Wap65). It encoded 439 amino acid residues with a signa l peptide of 22 residues and showed an amino acid sequence identity of 88% to that of goldfish reported before (J. Biol. Chem. 1995. 270: 17087-17092) . The number of potential N-linked glycosylation sites of carp Wap65 was tw o in contrast to three for goldfish. In addition, molecular mass determined by SDS-PAGE was apparently different from that of goldfish. These results suggest that the amount of oligosaccharide is different between the carp an d goldfish protein. As in goldfish, carp Wap65 mRNA showed marked accumulat ion in hepatopancreas of the 30 degreesC- acclimated fish, which was 8-fold higher than that of the 10 degreesC-acclimated fish. Carp Wap65 showed 30% amino acid identity to mammalian hemopexins, which appeared to be consider ably low in comparison with those among mammalian hemopexins (72 to 80%), o r among carp Wap65 and rainbow trout hemopexin-like protein (70%). However, although mammalian hemopexins contain residues comprising the heme binding pocket, carp Wap65 lacked one of the two histidine residues to serve as he me axial ligands in hemopexins. Our data on carp protein substantiates the previous observation for goldfish and indicates that Wap65 might have some important functions in warm-temperature-acclimation of fish.