Computational prediction of atomic structures of helical membrane proteins aided by EM maps

Biophys J. 2007 Sep 15;93(6):1950-9. doi: 10.1529/biophysj.106.102137. Epub 2007 May 11.

Abstract

Integral membrane proteins pose a major challenge for protein-structure prediction because only approximately 100 high-resolution structures are available currently, thereby impeding the development of rules or empirical potentials to predict the packing of transmembrane alpha-helices. However, when an intermediate-resolution electron microscopy (EM) map is available, it can be used to provide restraints which, in combination with a suitable computational protocol, make structure prediction feasible. In this work we present such a protocol, which proceeds in three stages: 1), generation of an ensemble of alpha-helices by flexible fitting into each of the density rods in the low-resolution EM map, spanning a range of rotational angles around the main helical axes and translational shifts along the density rods; 2), fast optimization of side chains and scoring of the resulting conformations; and 3), refinement of the lowest-scoring conformations with internal coordinate mechanics, by optimizing the van der Waals, electrostatics, hydrogen bonding, torsional, and solvation energy contributions. In addition, our method implements a penalty term through a so-called tethering map, derived from the EM map, which restrains the positions of the alpha-helices. The protocol was validated on three test cases: GpA, KcsA, and MscL.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Bacterial Proteins / chemistry
  • Bacterial Proteins / ultrastructure
  • Biophysical Phenomena
  • Biophysics
  • Glycophorins / chemistry
  • Glycophorins / ultrastructure
  • Humans
  • Hydrogen Bonding
  • Ion Channels / chemistry
  • Ion Channels / ultrastructure
  • Membrane Proteins / chemistry*
  • Membrane Proteins / ultrastructure*
  • Microscopy, Electron, Transmission
  • Models, Molecular*
  • Potassium Channels / chemistry
  • Potassium Channels / ultrastructure
  • Protein Structure, Secondary
  • Static Electricity
  • Thermodynamics

Substances

  • Bacterial Proteins
  • GYPA protein, human
  • Glycophorins
  • Ion Channels
  • Membrane Proteins
  • Potassium Channels
  • Tb-MscL protein, Mycobacterium tuberculosis
  • prokaryotic potassium channel