Structure-based function prediction of uncharacterized protein using binding sites comparison

PLoS Comput Biol. 2013;9(11):e1003341. doi: 10.1371/journal.pcbi.1003341. Epub 2013 Nov 14.

Abstract

A challenge in structural genomics is prediction of the function of uncharacterized proteins. When proteins cannot be related to other proteins of known activity, identification of function based on sequence or structural homology is impossible and in such cases it would be useful to assess structurally conserved binding sites in connection with the protein's function. In this paper, we propose the function of a protein of unknown activity, the Tm1631 protein from Thermotoga maritima, by comparing its predicted binding site to a library containing thousands of candidate structures. The comparison revealed numerous similarities with nucleotide binding sites including specifically, a DNA-binding site of endonuclease IV. We constructed a model of this Tm1631 protein with a DNA-ligand from the newly found similar binding site using ProBiS, and validated this model by molecular dynamics. The interactions predicted by the Tm1631-DNA model corresponded to those known to be important in endonuclease IV-DNA complex model and the corresponding binding free energies, calculated from these models were in close agreement. We thus propose that Tm1631 is a DNA binding enzyme with endonuclease activity that recognizes DNA lesions in which at least two consecutive nucleotides are unpaired. Our approach is general, and can be applied to any protein of unknown function. It might also be useful to guide experimental determination of function of uncharacterized proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / classification
  • Bacterial Proteins / metabolism*
  • Binding Sites*
  • Computational Biology / methods*
  • DNA / chemistry
  • DNA / metabolism
  • Models, Statistical
  • Molecular Dynamics Simulation
  • Protein Conformation
  • Thermotoga maritima

Substances

  • Bacterial Proteins
  • DNA

Grants and funding

Financial support was provided by grant P1-0002 of the Ministry of Higher Education, Science, and Technology of Slovenia and the Slovenian Research Agency. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.