Snake venom phospholipases A2. A fluorescence study of their binding to phospholipid vesicles correlation with their anticoagulant activities

• Published in: The Journal of biological chemistry 1980-08, Vol.255 (16), p.7734-7739
• Main Authors: Prigent-Dachary, J., Boffa, M.C., Boisseau, M.R., Dufourcq, J.
• Format: Article
• Language: English
• Subjects: Animals; Anticoagulants; Barium - pharmacology; Barium Compounds; Calcium - pharmacology; Chlorides - pharmacology; Crotalid Venoms - metabolism; Elapid Venoms - metabolism; Fluorescence; Hydrogen-Ion Concentration; Phosphatidylcholines - metabolism; Phospholipases - metabolism; Phospholipases A - metabolism; Phospholipases A2; Phospholipids - metabolism; Snake Venoms - metabolism; Snake Venoms - pharmacology; Viper Venoms - metabolism
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(19)43892-1

The interaction of snake venom phospholipases A2 with phospholipids has been studied by intrinsic fluorescence. This has been performed in order to understand why some enzymes possess anticoagulant properties while others have no action on blood clotting. We show that phospholipases A2 can be distinguished according to their binding properties to phospholipid vesicles. Strong inhibitors of coagulation interact with phospholipids with a significant change of their fluorescence while poor inhibitors have little or no effect. Strong inhibitors have a great affinity toward phosphatidylserine and do not require Ca2+ for interaction. Similar results are obtained with phosphatidylcholine-phosphatidylserine 1:1 mixtures. The diether analogue of phosphatidylcholine shows that formation of the complex is promoted by Ca2+ and can occur whenever the lipids are in crystal or fluid phase. Inactivation of anticoagulant phospholipase A2 decreased the affinity of enzyme for the phospholipids. The change in the intrinsic fluorescence of the phospholipases A2 on binding indicates a modification of the environment of the tryptophan residues. This is discussed in terms of the so-called interface recognition site as seen in the case of pancreatic phospholipase A2. It is concluded that the phospholipases may inhibit coagulation by competing with clotting proteins for the lipid surface. Although not considered in this study, the consequence of the hydrolysis of lipids remains to be estimated.