title: Binding site and affinity prediction of general anesthetics to protein targets using docking.
abstract: BACKGROUND
  The protein targets for general anesthetics remain unclear. A tool to predict anesthetic binding for potential binding targets is needed. In this study, we explored whether a computational method, AutoDock, could serve as such a tool.
  
  
  METHODS
  High-resolution crystal data of water-soluble proteins (cytochrome C, apoferritin, and human serum albumin), and a membrane protein (a pentameric ligand-gated ion channel from Gloeobacter violaceus [GLIC]) were used. Isothermal titration calorimetry (ITC) experiments were performed to determine anesthetic affinity in solution conditions for apoferritin. Docking calculations were performed using DockingServer with the Lamarckian genetic algorithm and the Solis and Wets local search method (http://www.dockingserver.com/web). Twenty general anesthetics were docked into apoferritin. The predicted binding constants were compared with those obtained from ITC experiments for potential correlations. In the case of apoferritin, details of the binding site and their interactions were compared with recent cocrystallization data. Docking calculations for 6 general anesthetics currently used in clinical settings (isoflurane, sevoflurane, desflurane, halothane, propofol, and etomidate) with known 50% effective concentration (EC(50)) values were also performed in all tested proteins. The binding constants derived from docking experiments were compared with known EC(50) values and octanol/water partition coefficients for the 6 general anesthetics.
  
  
  RESULTS
  All 20 general anesthetics docked unambiguously into the anesthetic binding site identified in the crystal structure of apoferritin. The binding constants for 20 anesthetics obtained from the docking calculations correlate significantly with those obtained from ITC experiments (P = 0.04). In the case of GLIC, the identified anesthetic binding sites in the crystal structure are among the docking predicted binding sites, but not the top ranked site. Docking calculations suggest a most probable binding site located in the extracellular domain of GLIC. The predicted affinities correlated significantly with the known EC(50) values for the 6 frequently used anesthetics in GLIC for the site identified in the experimental crystal data (P = 0.006). However, predicted affinities in apoferritin, human serum albumin, and cytochrome C did not correlate with these 6 anesthetics' known experimental EC(50) values. A weak correlation between the predicted affinities and the octanol/water partition coefficients was observed for the sites in GLIC.
  
  
  CONCLUSIONS
  We demonstrated that anesthetic binding sites and relative affinities can be predicted using docking calculations in an automatic docking server (AutoDock) for both water-soluble and membrane proteins. Correlation of predicted affinity and EC(50) for 6 frequently used general anesthetics was only observed in GLIC, a member of a protein family relevant to anesthetic mechanism.
authors.affiliation: Department of Anesthesiology and Critical Care, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. liur@uphs.upenn.edu
authors.forename: Renyu
authors.initials: R
authors.lastname: Liu
authors.forename: Jose Manuel
authors.initials: JM
authors.lastname: Perez-Aguilar
authors.forename: David
authors.initials: D
authors.lastname: Liang
authors.forename: Jeffery G
authors.initials: JG
authors.lastname: Saven
mesh_terms: D000465:Algorithms; D000777:Anesthetics / Q000097:blood / Q000493:pharmacokinetics*; D000818:Animals; D001052:Apoferritins / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D002151:Calorimetry; D045304:Cytochromes c / Q000737:chemistry / Q000378:metabolism; D006736:Horses; D006801:Humans; D007473:Ion Channels / Q000737:chemistry / Q000378:metabolism; D008024:Ligands; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D000442:Octanols / Q000737:chemistry; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D012709:Serum Albumin / Q000737:chemistry / Q000378:metabolism; D012995:Solubility; D013816:Thermodynamics; D014867:Water / Q000737:chemistry
chemical_list: D000777:Anesthetics; D007473:Ion Channels; D008024:Ligands; D000442:Octanols; D011506:Proteins; D012709:Serum Albumin; D014867:Water; D045304:Cytochromes c; D001052:Apoferritins