Regulation of vitamin K-dependent protein S. Inactivation by thrombin.
Thrombin treatment of the vitamin K-dependent protein S resulted in the loss of the activated protein C cofactor activity associated with protein S. The addition of phospholipid vesicles inhibited the inactivation. Thrombin treatment did not alter the molecular weight of the native protein. However, upon reduction, a peptide of approximately 3000 daltons was released from the treated protein. The interaction between calcium and protein S was reduced by thrombin treatment. When the calcium interaction was determined by the quenching of the intrinsic fluorescence of protein S, thrombin treatment appeared to inhibit the interaction between calcium and the protein. When the calcium interaction was observed by measuring the effect on the electrophoretic mobility of the protein, thrombin treatment reduced the interaction between calcium and protein S. However, the effect of thrombin treatment on the interaction between calcium and protein S was less than observed by the fluorescent method. This observation suggests that fluorescence quenching may be a result of a structural change induced by calcium binding. Thrombin treatment of protein S appears to uncouple the calcium binding from the structural change. In addition, the interaction between protein S and phospholipid vesicles was reduced by thrombin treatment. These results suggest that the thrombin conversion of protein S into a two-chain protein causes the loss of a calcium-induced change in protein structure, loss of the lipid-binding properties, and the loss of cofactor activity.
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D005165:Factor V / Q000378:metabolism; D015943:Factor Va; D006023:Glycoproteins / Q000037:antagonists & inhibitors*; D007700:Kinetics; D008970:Molecular Weight; D017293:Protein S; D013050:Spectrometry, Fluorescence; D013917:Thrombin / Q000378:metabolism*
D006023:Glycoproteins; D017293:Protein S; D015943:Factor Va; D005165:Factor V; D013917:Thrombin; D002118:Calcium