Interactions of the components of the prothrombinase complex.
The interaction of components of the prothrombinase complex, i.e. bovine Factor X or Factor Xa, bovine Factor V or Factor Va, phospholipid, and Ca2+, in various combinations was studied primarily by a gel filtration technique. In experiments, in which phospholipids ranging from those isolated from naturally occurring sources to those long chain (18 : 1) as well as short chain 6 : 0 and 7 : 0 fatty acids prepared by chemical and enzymatic synthesis were used, it was evident that a net negative surface charge on the lipid dispersions was one of the important requirements for interaction. Though the short chain fatty acid phospholipids interacted with the proteins of the prothrombinase complex, there was invariably a diminution in the activity of the enzyme complex. It was established that Factor V or Va did not bind Ca2+ and that the binding of either of these factors with phospholipids (with a net negative charge) was not dependent on Ca2+. However, the interaction of Factor X or Factor Xa with phospholipids with a negative charge required Ca2+. It was shown that Factor X could bind to the same type of lipid of lipid surface as that noted for Factor Xa. Of interest was the apparent difference in the phospholipid binding characteristics of the two variant forms of bovine plasma Factor X, i.e. X1 and X2, which might in part explain the differences in their specific activities. Of importance was the lack of demonstrable complex formation between Factors II, X and V in the absence of phospholipids and/or in the presence or absence of Ca2+. The significance of these results as they might apply to the configuration of the prothrombinase complex and its interaction with prothrombin plus  the usefulness of the short chain fatty acid phospholipid in exploring these lipid-protein interactions are discussed.
D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002417:Cattle; D002850:Chromatography, Gel; D005165:Factor V / Q000378:metabolism*; D005170:Factor X / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism*; D011485:Protein Binding; D011517:Prothrombin Time
D005227:Fatty Acids; D010446:Peptide Fragments; D010743:Phospholipids; D005165:Factor V; D005170:Factor X; D002118:Calcium