CCSV-MPase, a Novel Procoagulant Metalloproteinase from Cerastes cerastes Venom: Purification, Biochemical Characterization and Protein Identification

Abstract

A procoagulant metalloproteinase called CCSV-MPase was purified from C. cerastes venom by successive chromatographic methods starting with gel-filtration through Sephadex G-75; ion-exchange DEAE-Cellulose A-50; affinity chromatography on Benzamidine Sepharose 6B and RP-HPLC on a C8 column. CCSV-MPase has been isolated to an extent of about tenfolds and its molecular mass was evaluated at 70 kDa by SDS-PAGE. CCSV-MPase hydrolyzes casein and fibrinogene as natural substrates. Its proteolytic activity was inhibited by EDTA and 1.10-phenanthroline, a chelators of bivalent cation metals and Zn(2+) respectively. CCSV-MPase is therefore a Zn(2+)-metalloproteinase with fibrinogenolytic but not hemorrhagic activity. It greatly decreased levels of plasmatic fibrinogen when administered to rats for 24 h. This fibrinogenase hydrolyzes the Bβ chain of human fibrinogen in vitro releasing fibrinopeptide B only. LC MS/MS analysis of tryptic fragments of CCSV-MPase demonstrated that it showed some sequence similarities with four other venom metalloproteinases. CCSV-MPase could be considered as a potential therapeutic agent as it is a non-hemorrhagic enzyme and could be useful in thrombotic diseases because of its defibrinogenation of blood.

Grape seed extract neutralizes the effects of Cerastes cerastes cerastes post-synaptic neurotoxin in mouse diaphragm

“…Although many papers on the effect of C. cerastes venom fractions have been published (Daoud et al, 1986;Basheer et al, 1995;Laraba-Djebari et al, 1995;Marrakchi et al, 1995Marrakchi et al, , 1997Abu-Sinna et al, 2003;Boukhalfa-Abib et al, 2009;Chérifi et al, 2010), yet post-synaptic neurotoxins are among the poorly-studied venom fractions of vipers. To our knowledge, the current study reports for the first time the ultrastructural toxic effects produced in the diaphragm of sub-lethally intoxicated mice, by the post-synaptic neurotoxin isolated from the Egyptian sand viper, C. cerastes cerastes.…”

Isolated biomolecules of pharmacological interest in hemostasis from Cerastes cerastes venom

“…Therefore, the venom toxicity cannot be attributed to only one component [11]. However, most venom components produce beneficial effects when they act alone [12]. Snake venom also contains non-protein components including citrate, metal ions, carbohydrates, nucleotides as well as low concentrations of free amino acids and lipids [13-15].…”

“…In addition to their hydrolytic activity, PLA2s may display many other activities, such as edematous, neurotoxic, cardiotoxic, hemolytic, convulsive, antiplatelet, antitumoral and anticoagulant properties. Based on their anticoagulant activity, PLA2s can be clinically useful against thrombotic diseases and for the diagnosis and treatment of hemostatic disorders [17]. The anticoagulant activity is due to their ability to inhibit platelet aggregation through factor Xa (FXa) blockade.…”

“…However, similar to PLA2s, metalloproteinases could be clinically useful against thrombotic diseases thanks to their potential use in laboratory tests or as therapeutic agents [20,21]. These proteinases may be useful for investigating the mechanisms of blood coagulation and platelet aggregation [11,12]. …”

“…Most of these proteins act on blood coagulation, including PLA2, the thrombin-like enzymes RP34 and afaâcytin, anticoagulant protease fraction, aggregant serine protease, hemorrhagic metalloproteinase CcH1, and the non-hemorrhagic metalloproteinase CCSV-MPase [11,12,31-35]. In this paper, we report pharmacological activity of biomolecules isolated from Cerastes cerastes on hemostasis process.…”