Premium
Purification and characterization of a platelet aggregation inhibitor and anticoagulant Cc 5_NTase, CD 73‐like, from Cerastes cerastes venom
Author(s) -
Saoud Samah,
Chérifi Fatah,
Benhassine Traki,
LarabaDjebari Fatima
Publication year - 2017
Publication title -
journal of biochemical and molecular toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.526
H-Index - 58
eISSN - 1099-0461
pISSN - 1095-6670
DOI - 10.1002/jbt.21885
Subject(s) - chemistry , biochemistry , adenosine diphosphate , venom , iodoacetamide , apyrase , snake venom , platelet , enzyme , cysteine , biology , platelet aggregation , immunology
The present study is the first attempt to report the characterization of a nucleotidase from Cerastes cerastes venom. A 70 kDa 5′‐nucleotidase (Cc‐5′NTase) was purified to homogeneity. The amino acid sequence of Cc‐5′NTase displayed high homology with many nucleotidases. Its activity was optimal at pH 7 with a specific hydrolytic activity toward mono‐, di‐, and triphosphate adenylated nucleotides. Cc‐5′NTase preferentially hydrolyzed ADP and obeyed Michaelis–Menten kinetics. Among the metals and inhibitors tested, Ni 2+ and Mg 2+ completely potentiated enzyme activity, whereas EGTA, PMSF, iodoacetamide, vanillic acid, vanillyl mandelic acid, and 1,10‐phenanthroline partially abolished its activity. Cc‐5′NTase was not lethal for mice at 5 mg/kg and exhibited in vivo anticoagulant effect. It also dose‐dependently inhibited adenosine diphosphate‐induced platelet aggregation by converting adenosine diphosphate to adenosine and prohibited arachidonic acid‐induced aggregation but was not effective on fibrinogen‐induced aggregation. Cc‐5′NTase could be a good tool as pharmacological molecule in thrombosis diagnostic and/or therapy.