Conformational changes in human prothrombin as detected by antibody populations.

Abstract The amino-terminal peptides of human prothrombin corresponding to residues 1–51 and 52–156 have been isolated from a thrombin digest of prothrombin fragment 1. The products of digestion were purified by means of barium citrate and ammonium sulfate precipitations, followed by gel filtration and hydroxyapatite chromatographies. They were identified by their molecular sizes as well as their amino acid compositions. Peptides 1–51 (F 1 A) and 52–156 (F 1 B) were used as affinity ligands for the isolation of antibody populations from antisera that were elicited against human prothrombin or prothrombin fragment 1. These antibody populations displayed restricted specificity for the respective ligands as shown by competitive radioimmunoassays. They were used to study the conformational changes in prothrombin and fragment 1. The F 1 A-specific antibody populations detected a conformational change which is stabilized by calcium ions and which has a transition midpoint at ∼ 0.2 mM calcium ion concentration. The F 1 B-specific antibody populations identified a different conformational change which is destabilized by calcium ions and which has a transition midpoint at ∼ 0.5 mM calcium.