Interaction of blood coagulation factor Va with phospholipid vesicles examined by using lipophilic photoreagents.
Two different lipophilic photoreagents, [3H]adamantane diazirine and 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine (TID), have been utilized to examine the interactions of blood coagulation factor Va with calcium, prothrombin, factor Xa, and, in particular, phospholipid vesicles. With each of these structurally dissimilar reagents, the extent of photolabeling of factor Va was greater when the protein was bound to a membrane surface than when it was free in solution. Specifically, the covalent photoreaction with Vl, the smaller subunit of factor Va, was 2-fold higher in the presence of phosphatidylcholine/phosphatidylserine (PC/PS, 3:1) vesicles, to which factor Va binds, than in the presence of 100% PC vesicles, to which the protein does not bind. However, the magnitude of the PC/PS-dependent photolabeling was much less than has been observed previously with integral membrane proteins. It therefore appears that the binding of factor Va to the membrane surface exposes Vl to the lipid core of the bilayer, but that only a small portion of the Vl polypeptide is exposed to, or embedded in, the bilayer core. Addition of either prothrombin or active-site-blocked factor Xa to PC/PS-bound factor Va had little effect on the photolabeling of Vl with TID, but reduced substantially the covalent labeling of Vh, the larger subunit of factor Va. This indicates that prothrombin and factor Xa each cover nonpolar surfaces on Vh when the macromolecules associate on the PC/PS surface. It therefore seems likely that the formation of the prothrombinase complex involves a direct interaction between Vh and factor Xa and between Vh and prothrombin.(ABSTRACT TRUNCATED AT 250 WORDS)
D000218:Adamantane / Q000031:analogs & derivatives* / Q000378:metabolism; D000818:Animals; D001389:Azirines / Q000378:metabolism*; D002417:Cattle; D005165:Factor V / Q000378:metabolism*; D015943:Factor Va; D005170:Factor X / Q000378:metabolism; D015951:Factor Xa; D007457:Iodine Radioisotopes; D007700:Kinetics; D008081:Liposomes; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D010777:Photochemistry; D011485:Protein Binding; D011516:Prothrombin / Q000378:metabolism; D014316:Tritium
D001389:Azirines; D007457:Iodine Radioisotopes; D008081:Liposomes; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D014316:Tritium; C024080:adamantane diazirine; D015943:Factor Va; C033174:3-(trifluoromethyl)-3-(3-iodophenyl)diazirine; D005165:Factor V; D011516:Prothrombin; D005170:Factor X; D015951:Factor Xa; D000218:Adamantane