5'-Nucleotidase from Vipera lebetina venom

Toxicon. 2015 Jan:93:155-63. doi: 10.1016/j.toxicon.2014.11.234. Epub 2014 Nov 28.

Abstract

5'-Nucleotidase (5'-NT) is widely represented in animal tissues (CD73) as well as in almost all snake venoms. In the present study, a 5'-NT isoform has been isolated from Vipera lebetina venom. The homodimeric isoform consists of monomers with molecular masses of 60 kDa. The enzyme is thermolabile and has pH optimum at 7.5. The 5'-NT activity is inhibited by metal ions Fe(3+), Cu(2+) and Zn(2+), enhanced by Mn(2+) while Mg(2+) and Ca(2+) have no remarkable effect. In addition to 120-kDa protein there are higher molecular forms of 5'-NT present in the V. lebetina venom. The cloning and sequencing of the 5'-NT coding cDNA resulted in 5'-truncated construct. MALDI-TOF and Orbitrap mass-spectrometry of the tryptic peptides confirmed the translated N-terminally truncated protein sequence concordance to the 5'-NT isolated from the venom. The isolated protein strongly inhibited ADP- or collagen-induced platelet aggregation.

Keywords: 5′-nucleotidase; MALDI-TOF MS; Platelet aggregation; Snake venom; Vipera lebetina; cDNA cloning.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5'-Nucleotidase / genetics*
  • 5'-Nucleotidase / isolation & purification
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Chromatography, Gel
  • Chromatography, Ion Exchange
  • DNA Primers / genetics
  • Electrophoresis, Polyacrylamide Gel
  • Models, Molecular*
  • Molecular Sequence Data
  • Platelet Aggregation / drug effects
  • Protein Conformation
  • Sequence Analysis, DNA
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Viper Venoms / enzymology*
  • Viper Venoms / toxicity
  • Viperidae / genetics*
  • Viperidae / metabolism

Substances

  • DNA Primers
  • Viper Venoms
  • 5'-Nucleotidase

Associated data

  • GENBANK/AHJ80886
  • GENBANK/KF408296