Molecular cloning and identification of bottle-nosed dolphin p40(phox), p47(phox) and p67(phox).
The bottle-nosed dolphin NADPH oxidase cytosolic components, p40(phox), p47(phox) and p67(phox) 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 p40(phox), p47(phox) and p67(phox) clones contained open reading frames encoding predicted polypeptides of 339, 391 and 526 amino acids, respectively. Analysis of the p47(phox) and p67(phox) amino acid sequences showed two potential Src homology three domains and p40(phox) one. Comparison of the deduced amino acids showed that dolphin p40(phox) sequence shared 88.8% similarity with the human p40(phox), that dolphin p47(phox) sequence shared 87.7% similarity with the bovine p47(phox), and that dolphin p67(phox) shared 88.1% similarity with the bovine p67(phox). Western blot analysis using anti-human p40(phox), p47(phox) and p67(phox) antibodies demonstrated that dolphin neutrophil possesses p40(phox), p47(phox) and p67(phox) with similar molecular masses and structures, to each counterpart in human neutrophils, except for the p67(phox) COOH-terminus. These results suggest that dolphin NADPH oxidase cytosolic components have functional activities equivalent to those of human.
Inoue
Y
Y
Department of Preventive Veterinary Medicine and Animal Health, Nihon University School of Veterinary Medicine, Nihon University, 1866 Kameino, Fujisawa, 252-8510, Kanagawa, Japan.
Itou
T
T
Jimbo
T
T
Sakai
T
T
Ueda
K
K
Imajoh-Ohmi
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002417:Cattle; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004289:Dolphins / Q000097:blood / Q000235:genetics*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D019255:NADPH Oxidases / Q000737:chemistry / Q000235:genetics*; D009504:Neutrophils / Q000201:enzymology; D010750:Phosphoproteins / Q000235:genetics; D012313:RNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D019105:Random Amplified Polymorphic DNA Technique / Q000662:veterinary; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D018076:DNA, Complementary; D010750:Phosphoproteins; C085072:neutrophil cytosol factor 40K; C079789:neutrophil cytosol factor 67K; D012313:RNA; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1