Abstract
Human coagulation Factor X (FX), a member of the vitamin K-dependent serine protease family, is a crucial component of the human coagulation cascade. Activated FX (FXa) participates in forming the prothrombinase complex on activated platelets to convert prothrombin to thrombin in coagulation reactions. In the current study, 30-ns MD simulations were performed on both the open and closed states of human FXa. Root mean squares (RMS) fluctuations showed that structural fluctuations concentrated on the loop regions of FXa, and the presence of a ligand in the closed system resulted in larger fluctuations of the gating residues. The open system had a gating distance from 9.23 to 11.33 Å, i.e., significantly larger than that of the closed system (4.69–6.35 Å), which allows diversified substrates of variable size to enter. Although the solvent accessible surface areas (SASA) of FXa remained the same in both systems, the open system generally had a larger total SASA or hydrophobic SASA (or both) for residues surrounding the S4 pocket. Additionally, more hydrogen bonds were formed in the closed state than in the open state of FXa, which is believed to play a significant role in maintaining the closed confirmation of the aryl-binding site. Based on the results of MD simulations, we propose that an induced-fit mechanism governs the functioning of human coagulation FX, which helps provide a better understanding of the interactions between FXa and its substrate, and the mechanism of the conformational changes involved in human coagulation.
We’re sorry, something doesn't seem to be working properly.
Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.
References
Davie EW, Fujikawa K, Kisiel W (1991) The coagulation cascade: initiation, maintenance, and regulation. Biochemistry 30:10363–10370
Mann KG, Nesheim ME, Church WR, Haley P, Krishnaswamy S (1990) Surface-dependent reactions of the vitamin K-dependent enzyme complexes. Blood 76:1–16
Caldwell SH, Hoffman M, Lisman T, Macik BG, Northup PG, Reddy KR, Tripodi A, Sanyal AJ (2006) Coagulation disorders and hemostasis in liver disease: pathophysiology and critical assessment of current management. Hepatology 44:1039–1046
Kozek-Langenecker S (2007) Management of massive operative blood loss. Minerva Anestesiol 73:401–415
Soliman DE, Broadman LM (2006) Coagulation defects. Anesthesiol Clin 24:549–578, vii
Telfer TP, Denson KW, Wright DR (1956) A new coagulation defect. Br J Haematol 2:308–316
Graham JB, Barrow EM, Hougie C (1957) Stuart clotting defect. II. Genetic aspects of a new hemorrhagic state. J Clin Invest 36:497–503
Bode W, Mayr I, Baumann U, Huber R, Stone SR, Hofsteenge J (1989) The refined 1.9 A crystal structure of human alpha-thrombin: interaction with D-Phe-Pro-Arg chloromethylketone and significance of the Tyr-Pro-Pro-Trp insertion segment. EMBO J 8:3467–3475
Katz BA, Elrod K, Luong C, Rice MJ, Mackman RL, Sprengeler PA, Spencer J, Hataye J, Janc J, Link J, Litvak J, Rai R, Rice K, Sideris S, Verner E, Young W (2001) A novel serine protease inhibition motif involving a multi-centered short hydrogen bonding network at the active site. J Mol Biol 307:1451–1486
Bode W, Turk D, Karshikov A (1992) The refined 1.9-A X-ray crystal structure of d-Phe-Pro-Arg chloromethylketone-inhibited human alpha-thrombin: structure analysis, overall structure, electrostatic properties, detailed active-site geometry, and structure-function relationships. Protein Sci 1:426–471
Katz BA, Elrod K, Verner E, Mackman RL, Luong C, Shrader WD, Sendzik M, Spencer JR, Sprengeler PA, Kolesnikov A, Tai VW, Hui HC, Breitenbucher JG, Allen D, Janc JW (2003) Elaborate manifold of short hydrogen bond arrays mediating binding of active site-directed serine protease inhibitors. J Mol Biol 329:93–120
Katz BA, Mackman R, Luong C, Radika K, Martelli A, Sprengeler PA, Wang J, Chan H, Wong L (2000) Structural basis for selectivity of a small molecule, S1-binding, submicromolar inhibitor of urokinase-type plasminogen activator. Chem Biol 7:299–312
Katz BA, Spencer JR, Elrod K, Luong C, Mackman RL, Rice M, Sprengeler PA, Allen D, Janc J (2002) Contribution of multicentered short hydrogen bond arrays to potency of active site-directed serine protease inhibitors. J Am Chem Soc 124:11657–11668
Parker ET, Pohl J, Blackburn MN, Lollar P (1997) Subunit structure and function of porcine factor Xa-activated factor VIII. Biochemistry 36:9365–9373
Padmanabhan K, Padmanabhan KP, Tulinsky A, Park CH, Bode W, Huber R, Blankenship DT, Cardin AD, Kisiel W (1993) Structure of human des(1–45) factor Xa at 2.2 A resolution. J Mol Biol 232:947–966
Singh N, Briggs JM (2008) Molecular dynamics simulations of Factor Xa: insight into conformational transition of its binding subsites. Biopolymer 89:1104–1113
Wang JF, Wei DQ, Li L, Zheng SY, Li YX, Chou KC (2007) 3D structure modeling of cytochrome P450 2 C19 and its implication for personalized drug design. Biochem Biophys Res Commun 355:513–519
Wang JF, Wei DQ, Lin Y, Wang YH, Du HL, Li YX, Chou KC (2007) Insights from modeling the 3D structure of NAD(P)H-dependent D-xylose reductase of Pichia stipitis and its binding interactions with NAD and NADP. Biochem Biophys Res Commun 359:323–329
Wang JF, Wei DQ, Chen C, Li Y, Chou KC (2008) Molecular modeling of two CYP2C19 SNPs and its implications for personalized drug design. Protein Peptide Lett 15:27–32
Wang JF, Gong K, Wei DQ, Li YX, Chou KC (2009) Molecular dynamics studies on the interactions of PTP1B with inhibitors: from the first phosphate-binding site to the second one. Protein Eng Des Sel 22:349–355
Wang JF, Wei DQ, Chou KC (2009) Insights from investigating the interactions of adamantane-based drugs with the M2 proton channel from the H1N1 swine virus. Biochem Biophys Res Commun 388:413–417
Zeng QK, Du HL, Wang JF, Wei DQ, Wang XN, Li YX, Lin Y (2009) Reversal of coenzyme specificity and improvement of catalytic efficiency of Pichia stipitis xylose reductase by rational site-directed mutagenesis. Biotechnol Lett 31:1025–1029
Wang Y, Wei DQ, Wang JF (2010) Molecular dynamics studies on T1 lipase: insight into a double-flap mechanism. J Chem Inf Model 50:875–878
Guo X, Wang JF, Zhu Y, Wei DQ (2010) Recent progress on computer-aided inhibitor design of H5N1 influenza A virus. Curr Comput Aided Drug Des 6:139–146
Wang JF, Wei DQ (2009) Role of structural bioinformatics and traditional Chinese medicine databases in pharmacogenomics. Pharmacogenomics 10:1213–1215
Wang JF, Chou KC (2010) Molecular modeling of cytochrome P450 and drug metabolism. Curr Drug Metab 11:342–346
Wang JF, Chou KC (2010) Insights from studying the mutation-induced allostery in the M2 proton channel by molecular dynamics. Protein Eng Des Sel 23:663–666
Li L, Wei DQ, Wang JF, Chou KC (2007) Computational studies of the binding mechanism of calmodulin with chrysin. Biochem Biophys Res Commun 358:1102–1107
Wang JF, Wei DQ, Chou KC (2008) Drug candidates from traditional chinese medicines. Curr Top Med Chem 8:1656–1665
Lian P, Wei DQ, Wang JF, Chou KC (2011) An allosteric mechanism inferred from molecular dynamics simulations on phospholamban pentamer in lipid membranes. PLoS One 6:e18587
Wang JF, Wei DQ, Chou KC (2008) Pharmacogenomics and personalized use of drugs. Curr Top Med Chem 8:1573–1579
Gong K, Li L, Wang JF, Cheng F, Wei DQ, Chou KC (2009) Binding mechanism of H5N1 influenza virus neuraminidase with ligands and its implication for drug design. Med Chem 5:242–249
Wang JF, Chou KC (2011) Insights from modeling the 3D structure of New Delhi metallo-β-lactamse and its binding interactions with antibiotic drugs. PLoS One 6:e18414
Gu RX, Gu H, Xie ZY, Wang JF, Arias HR, Wei DQ, Chou KC (2009) Possible drug candidates for Alzheimer’s disease deduced from studying their binding interactions with alpha7 nicotinic acetylcholine receptor. Med Chem 5:250–262
Wang JF, Yan JY, Wei DQ, Chou KC (2009) Binding of CYP2C9 with diverse drugs and its implications for metabolic mechanism. Med Chem 5:263–270
Wang JF, Zhang CC, Chou KC, Wei DQ (2009) Structure of cytochrome p450s and personalized drug. Curr Med Chem 16:232–244
Chen Q, Zhang T, Wang JF, Wei DQ (2011) Advances in human cytochrome P450 and personalized medicine. Curr Drug Metab 12:436–444
Daura X, Haaksma E, van Gunsteren WF (2000) Factor Xa: simulation studies with an eye to inhibitor design. J Comput Aided Mol Des 14:507–529
Venkateswarlu D, Perera L, Darden T, Pedersen LG (2002) Structure and dynamics of zymogen human blood coagulation factor X. Biophys J 82:1190–1206
Corte JR, Fang T, Pinto DJ, Han W, Hu Z, Jiang XJ, Li YL, Gauuan JF, Hadden M, Orton D, Rendina AR, Luettgen JM, Wong PC, He K, Morin PE, Chang CH, Cheney DL, Knabb RM, Wexler RR, Lam PY (2008) Structure-activity relationships of anthranilamide-based factor Xa inhibitors containing piperidinone and pyridinone P4 moieties. Bioorg Med Chem Lett 18:2845–2849
Berman HM, Westbrook J, Feng Z, Gilliland G, Bhat TN, Weissig H, Shindyalov IN, Bourne PE (2000) The protein data bank. Nucleic Acids Res 28:235–242
Underwood MC, Zhong D, Mathur A, Heyduk T, Bajaj SP (2000) Thermodynamic linkage between the S1 stie, the Na + site, and the Ca2+ site in the protease domain of human coagulation factor xa. J Biol Chem 275:36876–36884
Griffon N, Di Stasio E (2001) Thermodynamics of Na + binding to coagulation serine proteases. Biophys Chem 90:89–96
Van Der Spoel D, Lindahl E, Hess B, Groenhof G, Mark AE, Berendsen HJ (2005) GROMACS: fast, flexible, and free. J Comput Chem 26:1701–1718
Scott WRP, Hüenenberger PH, Tironi IG, Mark AE, Billeter SR, Fennen J, Torda AE, Huber T, Krueger P, van Gunsteren WF (1999) The GROMOS biomolecular simulation program package. J Phys Chem A 103:3596–3607
Aalten DM van, Bywater R, Findlay JB, Hendlich M, Hooft RW, Vriend G (1996) PRODRG, a program for generating molecular topologies and unique molecular descriptors from coordinates of small molecules. J Comput Aided Mol Des 10:255–262
Matter H, Nazaré M, Güssregen S, Will DW, Schreuder H, Bauer A, Urmann M, Ritter K, Wagner M, Wehner V (2009) Evidence for C-Cl/C-Br…pi interactions as an important contribution to protein-ligand binding affinity. Angew Chem Int Edn Engl 48:2911–2916
Wallnoefer HG, Fox T, Liedl KR, Tautermann CS (2010) Dispersion dominate halogen-π interactions: energies and locations of minima. Phys Chem Chem Phys 12:14941–14949
Sreenivasan U, Axelsen PH (1992) Buried water in homologous serine proteases. Biochemistry 31:12785–12791
Guvench O, Price DJ, Brooks CL III (2005) Receptor rigidity and ligand mobility in trypsin-ligand complexes. Proteins 58:407–417
Lesk AM, Fordham WD (1996) Conservation and variability in the structures of serine proteinases of the chymotrypsin family. J Mol Biol 258:501–537
Wallnoefer HG, Handschuh S, Liedl KR, Fox T (2010) Stabilizing of a globular protein by a highly complex water network: a molecular dynamics simulation study on factor Xa. J Phys Chem B 114:7405–7412
Miller CA, Gellman SH, Abbott NL, de Pablo JJ (2009) Association of helical beta-peptides and their aggregation behavior from the potential of mean force in explicit solvent. Biophys J 96:4349–4362
Hlevnjak M, Zitkovic G, Zagrovic B (2010) Hydrophilicity matching: a potential prerequisite for the formation of protein-protein complexes in the cell. PLoS One 5:e11169
Lee C, Ham SJ (2011) Characterizing amyloid-beta protein misfolding from molecular dynamics simulations with explicit water. J Comput Chem 32:349–355
Acknowledgments
This work was supported by the grants from the National Basic Research Program of China (973 Program, No. 2011CB910204 and 2012CB316501), Research Program of CAS (KSCX2-EW-R-04), National Natural Science Foundation of China (No. 30900272), Shanghai Natural Science Foundation (No. 10ZR1421500), China Postdoctoral Science Foundation (No. 20110490068), and in part by Shanghai Pujiang Scholarship Program (No. 10PJ1408000). The authors gratefully acknowledge the support of SA-SIBS scholarship program.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wang, JF., Hao, P., Li, YX. et al. Exploration of conformational transition in the aryl-binding site of human FXa using molecular dynamics simulations. J Mol Model 18, 2717–2725 (2012). https://doi.org/10.1007/s00894-011-1295-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00894-011-1295-x