title: Purification and characterization of a platelet aggregation inhibitor and anticoagulant Cc 5_NTase, CD 73-like, from Cerastes cerastes venom.
abstract: 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, Ni2+and Mg2+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.
authors.affiliation: USTHB, Faculty of Biological Sciences, Laboratory of Cellular and Molecular Biology, BP 32 El-Alia, Bab Ezzouar, Algiers, Algeria.
authors.forename: Samah
authors.initials: S
authors.lastname: Saoud
authors.affiliation: USTHB, Faculty of Biological Sciences, Laboratory of Cellular and Molecular Biology, BP 32 El-Alia, Bab Ezzouar, Algiers, Algeria.
authors.forename: Fatah
authors.initials: F
authors.lastname: Chérifi
authors.affiliation: USTHB, Faculty of Biological Sciences, Laboratory of Cellular and Molecular Biology, BP 32 El-Alia, Bab Ezzouar, Algiers, Algeria.
authors.forename: Traki
authors.initials: T
authors.lastname: Benhassine
authors.affiliation: USTHB, Faculty of Biological Sciences, Laboratory of Cellular and Molecular Biology, BP 32 El-Alia, Bab Ezzouar, Algiers, Algeria.
authors.forename: Fatima
authors.initials: F
authors.lastname: Laraba-Djebari
mesh_terms: D015720:5'-Nucleotidase / Q000737:chemistry / Q000302:isolation & purification / Q000493:pharmacokinetics; D000818:Animals; D001792:Blood Platelets / Q000378:metabolism*; D006801:Humans; D010974:Platelet Aggregation / Q000187:drug effects*; D010975:Platelet Aggregation Inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D030162:Reptilian Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D014757:Viper Venoms / Q000737:chemistry*; D017819:Viperidae
chemical_list: D010975:Platelet Aggregation Inhibitors; D030162:Reptilian Proteins; D014757:Viper Venoms; D015720:5'-Nucleotidase
keywords: 5′-nucleotidases; ADPases; ATPases; coagulation; platelet aggregation