Molecular cloning and characterization of ecto-5'-nucleotidase from the venoms of Gloydius blomhoffi.
A Gloydius blomhoffi brevicaudus venom gland cDNA library was screened to isolate cDNA clones using probes based on highly conserved amino acid sequences from known ecto-5'-nucleotidases (ecto-5'-NTs). Molecular cloning of ecto-5'-NT from G. blomhoffi brevicaudus venom predicted that it was a glycosyl phosphatidylinositol-anchored membrane protein containing 588 amino acid residues with 7 potential N-linked glycosylation sites. The deduced amino acid sequence shows approximately 60% sequence identity to mammalian ecto-5'-NT sequences. This is the first report of the primary structure of ecto-5'-NT from a reptile. Gel-filtration chromatography of fresh venom from Gloydius blomhoffi blomhoffi, a subspecies of G. blomhoffi, revealed that at least a part of ecto-5'-NT is bound to exosome-like vesicles.
Ogawa
Yuko
Y
Institute of Medicinal Chemistry, Hoshi University, Ebara, Shinagawa-ku, Tokyo 142-8501, Japan.
Murayama
Nobuhiro
N
Yanoshita
Ryohei
R
D015720:5'-Nucleotidase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000818:Animals; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D003435:Crotalid Venoms / Q000737:chemistry / Q000201:enzymology*; D055354:Exosomes / Q000378:metabolism; D015723:Gene Library; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D017819:Viperidae / Q000378:metabolism*
D003435:Crotalid Venoms; D015720:5'-Nucleotidase