Structure determination using poorly diffracting membrane-protein crystals: the H+-ATPase and Na+,K+-ATPase case history

Acta Crystallogr D Biol Crystallogr. 2010 Mar;66(Pt 3):309-13. doi: 10.1107/S0907444909053244. Epub 2010 Feb 12.

Abstract

An approach is presented for the structure determination of membrane proteins on the basis of poorly diffracting crystals which exploits molecular replacement for heavy-atom site identification at 6-9 A maximum resolution and improvement of the heavy-atom-derived phases by multi-crystal averaging using quasi-isomorphous data sets. The multi-crystal averaging procedure allows real-space density averaging followed by phase combination between non-isomorphous native data sets to exploit crystal-to-crystal nonisomorphism despite the crystals belonging to the same space group. This approach has been used in the structure determination of H(+)-ATPase and Na(+),K(+)-ATPase using Ca(2+)-ATPase models and its successful application to the Mhp1 symporter using LeuT as a search model is demonstrated.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arabidopsis Proteins / chemistry*
  • Crystallography, X-Ray
  • Models, Molecular
  • Protein Structure, Tertiary
  • Proton-Translocating ATPases / chemistry*
  • Sodium-Potassium-Exchanging ATPase / chemistry*
  • Swine

Substances

  • Arabidopsis Proteins
  • Proton-Translocating ATPases
  • AHA2 protein, Arabidopsis
  • Sodium-Potassium-Exchanging ATPase