Metalloproteinase with factor X activating and fibrinogenolytic activities from Vipera berus berus venom

We have previously shown that Vipera berus berus venom contains several factor X activating enzymes. In the present study we have investigated one of them. The enzyme was separated from venom by gel filtration on Sephadex G-100 superfine and chromatography on agarose HPS-7 and phenyl-agarose. The en...

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Bibliographic Details
Published in:Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 2003-08, Vol.135 (4), p.575-582
Main Authors: Samel, Mari, Vija, Heiki, Subbi, Juhan, Siigur, Jüri
Format: Article
Language:English
Subjects:
Animals
Factor X - metabolism
Factor X activating enzyme
Fibrinogen - metabolism
Fibrinogenolytic activity
Glycoprotein
MALDI-TOF MS
Metalloproteases - isolation & purification
Metalloproteases - metabolism
Metalloproteinase
Peptides - metabolism
Snake venom
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Substrate Specificity
Viper Venoms - enzymology
Vipera berus berus
Viperidae - metabolism
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Summary:We have previously shown that Vipera berus berus venom contains several factor X activating enzymes. In the present study we have investigated one of them. The enzyme was separated from venom by gel filtration on Sephadex G-100 superfine and chromatography on agarose HPS-7 and phenyl-agarose. The enzyme is a glycosylated metalloproteinase containing hexoses, hexosamines and neuraminic acid. The purified factor X activating enzyme consists of two equal chains (59 kDa). The specificity studies have shown that enzyme is nonspecific factor X activating proteinase hydrolysing also proteins such as azocasein, gelatin and fibrinogen. The enzyme hydrolyses oxidized insulin B-chain at the positions Ala 14–Leu 15 and Tyr 16–Leu 17 but it is inactive on fibrin, plasminogen and prothrombin. We used 8–10 amino acid residues containing peptides, which reproduce the sequence around the cleavage sites in factor X, factor IX and fibrinogen, as potential substrates for enzyme. Cleavage products of peptide hydrolysis were determined by MALDI-TOF MS. The peptide Asn–Asn–Leu–Thr–Arg–Ile–Val–Gly–Gly—factor X fragment was cleaved by enzyme at positions Leu 3–Thr 4 and Arg 5–Ile 6. The fibrinogen peptide fragment Glu–Tyr–His–Thr–Glu–Lys–Leu–Val–Thr–Ser was hydrolysed at position Lys 6–Leu 7.
ISSN:1096-4959
1879-1107
DOI:10.1016/S1096-4959(03)00171-4