Interaction of an amphitropic protein (factor Xa) with membrane models in a complex system.
Phosphatidylserine (PS) plays a crucial role, in the conversion of prothrombin into thrombin by the protease, factor Xa. Physiologically, the conversion occurs in the prothrombinase complex. The question of how water-soluble proteins that normally circulate in plasma bind remains to be unambiguously determined. We previously found that the amphitropic proteins (prothrombin, factors V and Va) penetrate into phospholipid layers. AC polarography has allowed the detection for the first time of insertion of factor Xa into condensed monolayers containing phosphatidylserine (PS) and phosphatidylcholine (PC) either 100% PS or 25% PS in the presence of Ca2+. This observation demonstrates that part of factor Xa can cross the phospholipid polar headgroup/hydrocarbon chain interface. In parallel experiments, radioactive surface measurements permitted measuring binding of tritium-labeled factor Xa onto a PS monolayer and calculate an association constant, 6x10(6) M(-1). Penetration of factor Xa into PS-containing vesicles was investigated also using photoactivable 5-[125I]iodonaphthalene-1-azide, which binds selectively to the lipid embedded domains of the protein. These experiments suggest that Factor Xa penetrates preferentially by its heavy chain, an alternative mode of binding to the commonly accepted binding via its Gla domain. Interaction of factor Xa with PS vesicles also changes its apparent K(m) for S 2222.
Lecompte
M F
MF
INSERM U-466 and Department of Biochemistry IFR31, Toulouse Cedex 9, France. lecompte@cict.fr
D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism*; D004789:Enzyme Activation; D015951:Factor Xa / Q000378:metabolism*; D006801:Humans; D008954:Models, Biological; D010713:Phosphatidylcholines / Q000737:chemistry / Q000378:metabolism*; D010718:Phosphatidylserines / Q000737:chemistry / Q000378:metabolism*; D011516:Prothrombin / Q000737:chemistry / Q000378:metabolism
D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D011516:Prothrombin; D015951:Factor Xa