title: 113Cd NMR study of bovine prothrombin fragment 1 and factor X.
abstract: The interaction of Cd2+ with bovine prothrombin fragment 1, prothrombin intermediate 1, factor X, and a modified (Gla-domainless) factor X has been studied with 113Cd NMR. All the 113Cd resonances observed in this study were in the chemical shift range expected for oxygen ligands, suggesting that cadmium is binding at the same sites where calcium binds. Both fragment 1 and factor X displayed two major resonances, one near 10 ppm from 113Cd2+ that did not exchange rapidly with unbound 113Cd2+ (the high-affinity, or H, resonance) and one near -15 ppm from 113Cd2+ that exchanged rapidly with unbound 113Cd2+ (the low-affinity, or L, resonance). The difference between the chemical shift of the H resonance and the chemical shift range of -90 to -125 ppm that has been reported for three other small calcium-binding proteins is postulated to be due to different coordination geometries for monocarboxylate and dicarboxylate ligands; Cd2+ binds to fragment 1 and factor X through the dicarboxylate side chains of gamma-carboxyglutamate (Gla) residues. This allows contribution of only one oxygen per carboxyl group. At least one of the first few 113Cd2+ ions bound to fragment 1 did not appear in the 113Cd NMR spectrum until a total of five 113Cd2+ had been added. This could be due to exchange broadening of initial 113Cd2+ resonances due to sharing of ligands among several sites. Filling all sites would then restrict ligand exchange. Addition of Zn2+ displaced 113Cd2+ from the H resonance sites. Factor X did not display the interactions among ion binding sites proposed for fragment 1.
mesh_terms: D000818:Animals; D002104:Cadmium; D002417:Cattle; D005170:Factor X; D007554:Isotopes; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D010446:Peptide Fragments; D011487:Protein Conformation; D011498:Protein Precursors; D011516:Prothrombin; D015032:Zinc
chemical_list: D007554:Isotopes; D010446:Peptide Fragments; D011498:Protein Precursors; D002104:Cadmium; C029733:prothrombin fragment 1; D011516:Prothrombin; D005170:Factor X; D015032:Zinc