Molecular cloning and identification of bottle-nosed dolphin p40phox, p47phox and p67phox

The bottle-nosed dolphin NADPH oxidase cytosolic components, p40phox, p47phox and p67phox cDNA’s were cloned from mitogen stimulated peripheral white blood cell mRNA utilizing the reverse transcription-polymerase chain reaction. The sequences of these cDNAs showed that dolphin p40phox, p47phox and p67phox clones contained open reading frames encoding predicted polypeptides of 339, 391 and 526 amino acids, respectively. Analysis of the p47phox and p67phox amino acid sequences showed two potential Src homology three domains and p40phox one. Comparison of the deduced amino acids showed that dolphin p40phox sequence shared 88.8% similarity with the human p40phox, that dolphin p47phox sequence shared 87.7% similarity with the bovine p47phox, and that dolphin p67phox shared 88.1% similarity with the bovine p67phox. Western blot analysis using anti-human p40phox, p47phox and p67phox antibodies demonstrated that dolphin neutrophil possesses p40phox, p47phox and p67phox with similar molecular masses and structures, to each counterpart in human neutrophils, except for the p67phox COOH-terminus. These results suggest that dolphin NADPH oxidase cytosolic components have functional activities equivalent to those of human.