Virtual screening against highly charged active sites: identifying substrates of alpha-beta barrel enzymes

Biochemistry. 2005 Feb 15;44(6):2059-71. doi: 10.1021/bi0481186.

Abstract

We have developed a virtual ligand screening method designed to help assign enzymatic function for alpha-beta barrel proteins. We dock a library of approximately 19,000 known metabolites against the active site and attempt to identify the relevant substrate based on predicted relative binding free energies. These energies are computed using a physics-based energy function based on an all-atom force field (OPLS-AA) and a generalized Born implicit solvent model. We evaluate the ability of this method to identify the known substrates of several members of the enolase superfamily of enzymes, including both holo and apo structures (11 total). The active sites of these enzymes contain numerous charged groups (lysines, carboxylates, histidines, and one or more metal ions) and thus provide a challenge for most docking scoring functions, which treat electrostatics and solvation in a highly approximate manner. Using the physics-based scoring procedure, the known substrate is ranked within the top 6% of the database in all cases, and in 8 of 11 cases, it is ranked within the top 1%. Moreover, the top-ranked ligands are strongly enriched in compounds with high chemical similarity to the substrate (e.g., different substitution patterns on a similar scaffold). These results suggest that our method can be used, in conjunction with other information including genomic context and known metabolic pathways, to suggest possible substrates or classes of substrates for experimental testing. More broadly, the physics-based scoring method performs well on highly charged binding sites and is likely to be useful in inhibitor docking against polar binding sites as well. The method is fast (<1 min per ligand), due largely to an efficient minimization algorithm based on the truncated Newton method, and thus, it can be applied to thousands of ligands within a few hours on a small Linux cluster.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alanine Racemase / chemistry
  • Alanine Racemase / metabolism
  • Algorithms
  • Amino Acid Isomerases / chemistry
  • Amino Acid Isomerases / metabolism
  • Ammonia-Lyases / chemistry
  • Ammonia-Lyases / metabolism
  • Apoenzymes / chemistry
  • Apoenzymes / metabolism
  • Binding Sites
  • Carbon-Carbon Lyases / chemistry
  • Carbon-Carbon Lyases / metabolism
  • Computational Biology / methods*
  • Computational Biology / statistics & numerical data
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / metabolism
  • Holoenzymes / chemistry
  • Holoenzymes / metabolism
  • Intramolecular Lyases / chemistry
  • Intramolecular Lyases / metabolism
  • Ligands
  • Phosphopyruvate Hydratase / chemistry*
  • Phosphopyruvate Hydratase / metabolism*
  • Protein Structure, Secondary
  • Racemases and Epimerases / chemistry
  • Racemases and Epimerases / metabolism
  • Static Electricity
  • Substrate Specificity

Substances

  • Apoenzymes
  • Escherichia coli Proteins
  • Holoenzymes
  • Ligands
  • Carbon-Carbon Lyases
  • o-succinylbenzoic acid synthase
  • Phosphopyruvate Hydratase
  • Ammonia-Lyases
  • methylaspartate ammonia-lyase
  • Racemases and Epimerases
  • Amino Acid Isomerases
  • Alanine Racemase
  • glutamate racemase
  • mandelate racemase
  • Intramolecular Lyases
  • muconate cycloisomerase