Effect of membrane fluidity and fatty acid composition on the prothrombin-converting activity of phospholipid vesicles.
Vesicles composed of phospholipids with different fatty acyl side chains have been utilized to examine the importance of the nonpolar membrane region for the prothrombin-converting activity of procoagulant phospholipid vesicles. Membranes composed of phosphatidylserine (PS) and phosphatidylcholine (PC) with unsaturated fatty acyl side chains were more active in prothrombin activation than membranes composed of phospholipids with saturated fatty acyl chains. This phenomenon was observed above the phase transition temperature, i.e., on membranes in the liquid-crystalline state. The prothrombin-converting activity of saturated phospholipids approached the activity of unsaturated phospholipids at high factor Va concentrations, which is indicative for a less favorable equilibrium constant for prothrombinase assembly on membrane surfaces composed of saturated phospholipids. The difference between saturated and unsaturated phospholipids was annulled on membranes with high mole percentages of PS. This may result from a compensating contribution of electrostatic forces to the binding equilibria involved in prothrombinase assembly. Additional effects on the prothrombin-converting activity were observed when membranes containing saturated phospholipids were studied below their phase transition temperature. In agreement with Higgins et al. [(1985) J. Biol. Chem. 260, 3604-3612], we found that the time required for the assembly of prothrombinase from membrane-bound factors Xa and Va is considerably prolonged on solid membranes. However, we also observed an effect of membrane fluidity on the steady-state rate of prothrombin activation. Kinetic experiments at saturating factor Va concentrations showed that the transition from the liquid-crystalline to the gel state caused a more than 9-fold decrease of the kcat of prothrombin activation without affecting the Km for prothrombin.(ABSTRACT TRUNCATED AT 250 WORDS)
Govers-Riemslag
J W
JW
Department of Biochemistry, University of Limburg, Maastricht, The Netherlands.
Janssen
M P
MP
Zwaal
R F
RF
Rosing
J
J
D000818:Animals; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D015943:Factor Va / Q000378:metabolism; D015951:Factor Xa / Q000378:metabolism; D005227:Fatty Acids / Q000032:analysis*; D005231:Fatty Acids, Unsaturated / Q000032:analysis; D007700:Kinetics; D008081:Liposomes / Q000737:chemistry* / Q000378:metabolism*; D008560:Membrane Fluidity / Q000502:physiology*; D008563:Membrane Lipids / Q000737:chemistry / Q000502:physiology; D010713:Phosphatidylcholines / Q000032:analysis / Q000378:metabolism; D010718:Phosphatidylserines / Q000032:analysis / Q000378:metabolism; D011516:Prothrombin / Q000378:metabolism*; D013696:Temperature; D013925:Thromboplastin / Q000378:metabolism
D005227:Fatty Acids; D005231:Fatty Acids, Unsaturated; D008081:Liposomes; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D015943:Factor Va; D011516:Prothrombin; D013925:Thromboplastin; D015951:Factor Xa