title: Solution conformations of the gamma-carboxyglutamic acid domain of bovine prothrombin fragment 1, residues 1-65.
abstract: Molecular dynamics simulations have been performed (AMBER version 3.1) on solvated residues 1-65 of bovine prothrombin fragment 1 (BF1) by using the 2.8-A resolution crystallographic coordinates as the starting conformation for understanding calcium ion-induced conformational changes that precede experimentally observable phospholipid binding. Simulations were performed on the non-metal-bound crystal structure, the form resulting from addition of eight calcium ions to the 1-65 region of the crystal structure, the form resulting from removal of calcium ions after 107 ps and continuing the simulation, and an isolated hexapeptide loop (residues 18-23). In all cases, the 100-ps time scale seemed adequate to sample an ensemble of solution conformers within a particular region of conformation space. The non-metal-containing BF1 did not unfold appreciably during a 106-ps simulation starting from the crystallographic geometry. The calcium ion-containing structure (Ca-BF1) underwent an interesting conformational reorganization during its evolution from the crystal structure: during the time course of a 107-ps simulation, Ca-BF1 experienced a trans----cis isomerization of the gamma-carboxyglutamic acid-21 (Gla-21)-Pro-22 peptide bond. Removal of the calcium ions from this structure followed by 114 ps of additional molecular dynamics showed significant unfolding relative to the final 20-ps average structure of the 107-ps simulation; however, the Gla-21-Pro-22 peptide bond remained cis. A 265-ps simulation on the termini-protected hexapeptide loop (Cys-18 to Cys-23) containing two calcium ions also did not undergo a trans----cis isomerization. It is believed that the necessary activation energy for the transitional event observed in the Ca-BF1 simulation was largely supplied by global conformational events with a possible assist from relief of intermolecular crystal packing forces. The presence of a Gla preceding Pro-22, the inclusion of Pro-22 in a highly strained loop structure, and the formation of two long-lived salt bridges prior to isomerization may all contribute to this finding.
authors.affiliation: Chemistry Department, University of North Carolina, Chapel Hill 27599-3290.
authors.forename: P S
authors.initials: PS
authors.lastname: Charifson
authors.forename: T
authors.initials: T
authors.lastname: Darden
authors.forename: A
authors.initials: A
authors.lastname: Tulinsky
authors.forename: J L
authors.initials: JL
authors.lastname: Hughey
authors.forename: R G
authors.initials: RG
authors.lastname: Hiskey
authors.forename: L G
authors.initials: LG
authors.lastname: Pedersen
mesh_terms: D015055:1-Carboxyglutamic Acid; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D002151:Calorimetry; D002417:Cattle; D003198:Computer Simulation; D008958:Models, Molecular; D010446:Peptide Fragments / Q000737:chemistry; D011487:Protein Conformation; D011516:Prothrombin / Q000737:chemistry* / Q000378:metabolism; D012996:Solutions
chemical_list: D010446:Peptide Fragments; D012996:Solutions; D015055:1-Carboxyglutamic Acid; D011516:Prothrombin; D002118:Calcium