Biochemical characterization of basilase, a fibrinolytic enzyme from Crotalus basiliscus basiliscus.
Snake venoms, especially from the Crotalidae family, contain a variety of enzymes that prevent blood coagulation by virtue of their fibrinolytic enzymes. Nineteen snake venoms were screened for fibrinolytic activity and the highest activity was found in the venom of Crotalus basiliscus basiliscus venom. The active principle, basilase, was isolated, purified, and found to have fibrinolytic and fibrinogenolytic activity. It had a molecular weight of 22,000 and 1 mol of zinc per mole of protein associated with it. The proteolytic activity of the enzyme against dimethyl casein was inhibited by ethylenediaminetetraacetic acid and alpha 2-macroglobulin. It did not inactivate alpha 2-macroglobulin. Basilase did not have any of the following activities: thrombin-like, factor X-like, protein C activating, or urokinase-like. It caused neither hemorrhage nor platelet aggregation. In spite of its proteolytic activity, basilase did not hydrolyze the membranes of platelets. Basilase had 24% alpha-helix, 31% beta-sheet, 25% turns, and 20% unordered structure, as determined by Fourier Transform Infrared spectroscopy. Basilase is an enzyme that hydrolyzes fibrin directly without activation of plasminogen.
Datta
G
G
Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins 80523.
Dong
A
A
Witt
J
J
Tu
A T
AT
D000595:Amino Acid Sequence; D002118:Calcium / Q000032:analysis; D002863:Chromogenic Compounds; D003435:Crotalid Venoms / Q000737:chemistry*; D006868:Hydrolysis; D008666:Metalloendopeptidases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010974:Platelet Aggregation; D017433:Protein Structure, Secondary; D017385:Sequence Homology; D012964:Sodium / Q000032:analysis; D017550:Spectroscopy, Fourier Transform Infrared; D013379:Substrate Specificity; D015032:Zinc / Q000032:analysis
D002863:Chromogenic Compounds; D003435:Crotalid Venoms; D012964:Sodium; D008666:Metalloendopeptidases; C089315:basiliscusfibrase; D015032:Zinc; D002118:Calcium