Characterization and cDNA cloning of aminopeptidase A from the venom of Gloydius blomhoffi brevicaudus.
The aminopeptidase activities of snake venoms from Gloydius blomhoffi brevicaudus, Gloydius halys blomhoffii, Trimeresurus flavoviridis, Bothrops jararaca and Crotalus atrox were investigated. Aminopeptidase A (APA), aminopeptidase B and aminopeptidase N activities were present in all snake venoms. The strongest APA activity was found in venom from G. blomhoffi brevicaudus. The susceptibility to metallopeptidase inhibitors and the pH optimum of the partially purified enzyme from G. blomhoffi brevicaudus venom were similar to those of known APAs from mammals. A G. blomhoffi brevicaudus venom gland cDNA library was screened to isolate cDNA clones using probes based on highly conserved amino acid sequences in known APAs. Molecular cloning of APA from G. blomhoffi brevicaudus venom predicted that it was a type II integral membrane protein containing 958 amino acid residues with 17 potential N-linked glycosylation sites. It possessed a His-Glu-Xaa-Xaa-His-(Xaa)(18)-Glu zinc binding motif that allowed the classification of this protein as a member of the M1 family of zinc-metallopeptidases, or gluzincins. The deduced amino acid sequence shows approximately 60% sequence identity to mammalian APA sequences. This is the first study to report the primary structure of APA from a reptile.
Ogawa
Yuko
Y
Institute of Medicinal Chemistry, Hoshi University, Shinagawa-ku, Tokyo 142-8501, Japan.
Murayama
Nobuhiro
N
Fujita
Yoshiaki
Y
Yanoshita
Ryohei
R
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D016000:Cluster Analysis; D003435:Crotalid Venoms / Q000201:enzymology*; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D043384:Glutamyl Aminopeptidase / Q000032:analysis / Q000235:genetics*; D008565:Membrane Proteins / Q000032:analysis / Q000235:genetics*; D008969:Molecular Sequence Data; D010802:Phylogeny; D011487:Protein Conformation; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D017819:Viperidae
D003435:Crotalid Venoms; D017931:DNA Primers; D018076:DNA, Complementary; D008565:Membrane Proteins; D043384:Glutamyl Aminopeptidase