title: Identification of a major enzyme for the synthesis and hydrolysis of cyclic ADP-ribose in amphibian cells and evolutional conservation of the enzyme from human to invertebrate.
abstract: Cyclic ADP-ribose (cADPR), a metabolite of NAD(+), is known to function as a second messenger for intracellular Ca(2+) mobilization in various vertebrate and invertebrate tissues. In this study, we isolated two Xenopus laevis cDNAs (frog cd38 and cd157 cDNAs) homologous to the one encoding the human cADPR-metabolizing enzyme CD38. Frog CD38 and CD157 are 298-amino acid proteins with 35.9 and 27.2 % identity to human CD38 and CD157, respectively. Transfection of expression vectors for frog CD38 and CD157 into COS-7 cells revealed that frog CD38 had NAD(+) glycohydrolase, ADP-ribosyl cyclase (ARC), and cADPR hydrolase activities, and that frog CD157 had no enzymatic activity under physiological conditions. In addition, when recombinant CD38 and frog brain homogenate were electrophoresed on an SDS-polyacrylamide gel, ARC of the brain homogenate migrated to the same position in the gel as that of frog CD38, suggesting that frog CD38 is the major enzyme responsible for cADPR metabolism in amphibian cells. The frog cd38 gene consists of eight exons and is ubiquitously expressed in various tissues. These findings provide evidence for the existence of the CD38-cADPR signaling system in frog cells and suggest that the CD38-cADPR signaling system is conserved during vertebrate evolution.
authors.affiliation: Department of Biochemistry, Kanazawa Medical University School of Medicine, 1-1 Daigaku, Uchinada, Kahoku-gun, Ishikawa 920-0293, Japan. tikeda@kanazawa-med.ac.jp
authors.forename: Takayuki
authors.initials: T
authors.lastname: Ikeda
authors.forename: Shin
authors.initials: S
authors.lastname: Takasawa
authors.forename: Naoya
authors.initials: N
authors.lastname: Noguchi
authors.forename: Koji
authors.initials: K
authors.lastname: Nata
authors.forename: Akiyo
authors.initials: A
authors.lastname: Yamauchi
authors.forename: Iwao
authors.initials: I
authors.lastname: Takahashi
authors.forename: Takeo
authors.initials: T
authors.lastname: Yoshikawa
authors.forename: Akira
authors.initials: A
authors.lastname: Sugawara
authors.forename: Hideto
authors.initials: H
authors.lastname: Yonekura
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Okamoto
mesh_terms: D036541:ADP-ribosyl Cyclase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D051997:ADP-ribosyl Cyclase 1 / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D001921:Brain / Q000201:enzymology; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D017124:Conserved Sequence; D036563:Cyclic ADP-Ribose / Q000096:biosynthesis* / Q000378:metabolism; D019143:Evolution, Molecular; D058851:GPI-Linked Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D006801:Humans; D006868:Hydrolysis; D007292:Inosine Nucleotides / Q000737:chemistry; D007700:Kinetics; D008969:Molecular Sequence Data; D009243:NAD / Q000031:analogs & derivatives / Q000737:chemistry; D009928:Organ Specificity; D010802:Phylogeny; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D017422:Sequence Analysis, DNA; D029867:Xenopus Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D014982:Xenopus laevis / Q000235:genetics*
chemical_list: D015703:Antigens, CD; D058851:GPI-Linked Proteins; D007292:Inosine Nucleotides; D011993:Recombinant Fusion Proteins; D029867:Xenopus Proteins; C107267:cyclic IDP-ribose; D009243:NAD; D036563:Cyclic ADP-Ribose; C032140:nicotinamide-hypoxanthine dinucleotide; D036541:ADP-ribosyl Cyclase; C087521:ADP-ribosyl cyclase 2; D051997:ADP-ribosyl Cyclase 1