title: Interaction of prothrombin with a phospholipid surface: evidence for a membrane-induced conformational change.
abstract: Prothrombin interacts with phosphatidylserine containing platelet membranes via its N-terminal, γ-carboxyglutamate (gla) residue-rich domain. Once bound it is cleaved to form the active protease, thrombin (factor IIa). Human prothrombin was cleaved with cathepsin G in the absence of calcium and magnesium ions. Under these conditions, the gla domain was removed. Phospholipid protected the protein from this proteolytic event, and this suggests that a conformational change may be induced by interaction with phospholipids. Binding of prothrombin to a surface containing 20% phosphatidylserine/80% phosphatidylcholine was detected by surface plasmon resonance, whereas no interaction with gla-domainless prothrombin was observed. Binding of intact prothrombin in the presence of calcium ions showed complex association kinetics, suggesting multiple modes of initial interaction with the surface. The kinetics of the dissociation phase could be fitted to a two-phase, exponential decay. This implies that there are at least two forms of the protein on the surface one of which dissociates tenfold more slowly than the other. Taken together, these data suggest that, on binding to a membrane surface, prothrombin undergoes a conformational change to a form which binds more tightly to the membrane.
authors.affiliation: School of Biological Sciences, Queen's University Belfast, Medical Biology Centre, Belfast, UK.
authors.forename: David F
authors.initials: DF
authors.lastname: Houston
authors.forename: David J
authors.initials: DJ
authors.lastname: Timson
mesh_terms: D015055:1-Carboxyglutamic Acid / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D056649:Cathepsin G / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008051:Lipid Bilayers; D008954:Models, Biological; D008968:Molecular Conformation; D010713:Phosphatidylcholines / Q000737:chemistry / Q000378:metabolism*; D010718:Phosphatidylserines / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D011499:Protein Processing, Post-Translational; D011516:Prothrombin / Q000737:chemistry / Q000378:metabolism*; D020349:Surface Plasmon Resonance; D013917:Thrombin / Q000378:metabolism*
chemical_list: D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D015055:1-Carboxyglutamic Acid; D011516:Prothrombin; C534094:CTSG protein, human; D056649:Cathepsin G; D013917:Thrombin; D002118:Calcium