Circular dichroism analysis of the secondary structures of bovine blood coagulation factor IX, factor X, and prothrombin.
Analysis of the far-ultraviolet circular dichroism spectrum of bovine blood coagulation factor IX reveals the presence of approximately 14% helical structures 26% beta-sheets, 20% beta-turns, and 40% coils. These values are essentially the same for the activation products of this zymogen, factor IX alpha alpha and factor IX alpha beta. Similar analysis for bovine factor X permits calculation of these secondary structural as approximately 11% helices, 31% beta-structures, 22% beta-turns, and 36% random structures. Bovine prothrombin contains approximately 12% helical structures, 35% beta-structures, 24% beta-turns, and 29% coils. None of these values is substantially altered as a result of increase of the pH from 7.4 to 10.5, or upon addition of Ca2+ to a concentration of at least 20 mM. Analysis of the near-ultraviolet spectra of factor IX and prothrombin suggests that several aromatic amino acid residues and the disulfide bond present in their gamma-carboxyglutamic acid-containing regions are exposed to solvent and are perturbed by the above pH adjustment and Ca2+ addition. Similar effects are observed in the case of factor X; in addition, the Trp residue at the amino terminus of the heavy chain appears to be influenced by the above pH alteration. The results reported in this paper show that these vitamin K-dependent blood coagulation proteins are similar in their ordered secondary structures, which are dominated by beta-sheets and beta-turns. Their overall secondary structures are not influenced by Ca2+ binding and are stable to alkaline pH changes. However, these same environmental alterations appear to be effective probes of aromatic residues in the gamma-carboxyglutamic acid regions.
Beals
J M
JM
Department of Chemistry, University of Notre Dame, Indiana 46556.
Castellino
F J
FJ
D000818:Animals; D002417:Cattle; D002942:Circular Dichroism; D005164:Factor IX; D005170:Factor X; D011487:Protein Conformation; D011516:Prothrombin; D013056:Spectrophotometry, Ultraviolet
D011516:Prothrombin; D005164:Factor IX; D005170:Factor X