Crystallography of the integral membrane protein EmrE from Escherichia coli

Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 2):2399-402. doi: 10.1107/S090744490402548X. Epub 2004 Nov 26.

Abstract

Crystals of the EmrE membrane-protein imposed several technical challenges for X-ray crystallography, including high mosaicity, poor diffraction and a relatively large number of heavy atoms. Consequently, the heavy-atom substructure solution was difficult to obtain. By removing the histidine tag for protein purification, the mosaicity and the diffraction quality were greatly improved. The direct-methods Shake-and-Bake program SnB was successful in locating the heavy-atom sites from a mutant of EmrE which lacks a cysteine and therefore has a reduction in the number of heavy-atom sites. The substructure solution was solved from data with anomalous difference at a resolution of 5.5 A and the structure was determined to 3.8 A.

Publication types

  • 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

  • Antiporters / chemistry*
  • Crystallography, X-Ray
  • Cysteine / chemistry
  • Drug Resistance, Multiple
  • Escherichia coli / metabolism*
  • Escherichia coli Proteins
  • Histidine / chemistry
  • Membrane Proteins / chemistry*
  • Models, Molecular
  • Mutation
  • Polymerase Chain Reaction
  • Protein Conformation

Substances

  • Antiporters
  • Escherichia coli Proteins
  • Membrane Proteins
  • EmrE protein, E coli
  • Histidine
  • Cysteine