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Purification and properties of the main coagulant and anticoagulant principles of Vipera Russell II snake venom

Authors
Journal
Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology
0167-4838
Publisher
Elsevier
Publication Date
Volume
786
Issue
3
Identifiers
DOI: 10.1016/0167-4838(84)90090-6
Keywords
  • Snake Venom
  • Coagulation
  • Phospholipase
  • Procoagulant
  • Anticoagulant
  • (V Russellii)
Disciplines
  • Biology

Abstract

Abstract Vipera russellii venom was separated into thirteen fractions by means of DEAE-Sephadex A-50 column chromatography. Fraction III possessed anticoagulant and phospholipase A activities and Fraction XI possessed procoagulant and caseinolytic activities, both were further purified by gel filtration on Sephacryl S-200 column. Purified procoagulant (Component II) was a two-chain protein with molecular weight of 86 000 consistng of A-chain ( M r 66 000) and B-chain ( M r 20 000). It was a glycoprotein containing 7.8% neutral sugar and 715 amino-acid residues. The procoagulant activity was 10-times that of the crude venom. It was an acidic proteinase with isoelectric point of pH 4.2. Upon heat treatment at 60°C, Component II was stable at pH 5.5. and 7.2 for 3 h, but was destroyed completely after 30 min at pH 8.9. It was devoid of esterase or amidase activity. Purified anticoagulant (Component I) was a single peptide chain with molecular weight of 16 000. It was carbohydrate free and contained 136 amino-acid residues. It was a basic protein with an isoelectric point of larger than pH 10. It was a potent phospholipase A with an enzymatic activity of 510 ± 30 μmol/min per mg using phosphatidylcholine as substrate, and 1 μg/ml was sufficient to cause 100% hemolysis by the indirect hemolytic method. Upon heat treatment at 90°C. Component I was heat stabe at pH 5.5. for more than 3 h, but was destroyed completely after 2 h at pH 7.2 and 8.9. The anticoagulant activity of Component I could be neutralized by platelet factor 3, tissue thromboplastin and cephalin.

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