Afaâcytin, an alpha beta-fibrinogenase from Cerastes cerastes (horned viper) venom, activates purified factor X and induces serotonin release from human blood platelets.
Afaâcytin, a proteinase with caseinolytic, arginine-esterase and amidase activities, was purified from the venom of Cerastes cerastes (horned viper) in two steps by gel filtration through Sephadex G75, then HPLC on carboxymethyl-cellulose. Afaâcytin has an isoelectric point of 6.25, and consists of two subunits, alpha and beta, which have the same apparent molecular mass (40,000) and are indistinguishable in the absence of reduction or/and deglycosylation. Subunit beta is constituted of two disulfide-linked polypeptidic chains, beta and beta'. The respective apparent molecular mass of the chains are 43,000 (alpha), 35,500 (beta) and 10,200 (beta') as determined by SDS/PAGE under reducing conditions. Both chains alpha and beta are N-glycosylated. The two chains have the same N-terminal sequence (20 residues) which is similar to those of other proteinases from snake venom. Susceptibility of afaâcytin to diisopropyl fluorophosphate and benzamidine indicates the presence of a serine and an aspartic (or glutamic) acid residues in the catalytic site. Ca2+ appears to be required for structural cohesion of the afaâcytin molecule. Afaâcytin exhibits alpha beta-fibrinogenase and alpha-fibrinase properties. It replaces missing factors VIII and IX in deficient plasmas, and activates purified human factor X into factor Xa. It releases serotonin from platelets and directly aggregates human (but not rabbit) blood platelets. Despite its thrombin-like characteristics, however, afaâcytin is not inhibited by plasmatic thrombin inhibitors. The procoagulant properties of afaâcytin therefore have potential clinical applications.
Centre National de la Recherche Scientifique, Institut Fédératif de Recherche Jean Roche, Université d'Aix-Marseille II, France.
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F
Laraba-Djebari
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MF
Martin-Eauclaire
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Mauco
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Marchot
D000595:Amino Acid Sequence; D000818:Animals; D001792:Blood Platelets / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D005170:Factor X / Q000378:metabolism*; D005340:Fibrinogen / Q000378:metabolism*; D006801:Humans; D008666:Metalloendopeptidases / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D008969:Molecular Sequence Data; D015539:Platelet Activation / Q000187:drug effects; D010974:Platelet Aggregation / Q000187:drug effects; D011817:Rabbits; D016415:Sequence Alignment; D012697:Serine Endopeptidases / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D012701:Serotonin / Q000378:metabolism*; D014757:Viper Venoms / Q000201:enzymology*; D017819:Viperidae / Q000378:metabolism*
D014757:Viper Venoms; D012701:Serotonin; D005170:Factor X; D005340:Fibrinogen; D012697:Serine Endopeptidases; C097119:afaacytin; D008666:Metalloendopeptidases