title: Comparison of the membrane binding kinetics of bovine prothrombin and its fragment 1.
abstract: Total internal reflection fluorescence microscopy has been used to compare the membrane binding characteristics of fluorescein-labeled bovine prothrombin and fluorescein-labeled bovine prothrombin fragment 1. The Ca(2+)-dependent association of these proteins with quartz-supported planar membranes composed of mixtures of phosphatidylserine (2-10 mol%) and phosphatidylcholine was examined. Equilibrium binding measurements showed that the apparent equilibrium dissociation constants increased with decreasing molar fractions of phosphatidylserine and that the dissociation constants were somewhat lower for intact prothrombin. Kinetic measurements, using fluorescence photobleaching recovery, showed that the measured dissociation rates were approximately equivalent for prothrombin and fragment 1 and did not change with the protein solution concentration or the molar fraction of phosphatidylserine. The kinetic data also implied that the surface binding mechanism for both proteins is more complex than a simple reversible reaction between monovalent proteins and monovalent surface sites. Measured equilibrium and kinetic constants are reported and compared for prothrombin and fragment 1 on planar membranes.
authors.affiliation: Department of Chemistry, University of North Carolina, Chapel Hill 27599.
authors.forename: K H
authors.initials: KH
authors.lastname: Pearce
authors.forename: M
authors.initials: M
authors.lastname: Hof
authors.forename: B R
authors.initials: BR
authors.lastname: Lentz
authors.forename: N L
authors.initials: NL
authors.lastname: Thompson
mesh_terms: D000818:Animals; D002417:Cattle; D066298:In Vitro Techniques; D007700:Kinetics; D008567:Membranes, Artificial; D008856:Microscopy, Fluorescence; D010446:Peptide Fragments / Q000378:metabolism; D010713:Phosphatidylcholines / Q000737:chemistry; D010718:Phosphatidylserines / Q000737:chemistry; D011516:Prothrombin / Q000378:metabolism*
chemical_list: D008567:Membranes, Artificial; D010446:Peptide Fragments; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D011516:Prothrombin