NMR investigation of the multidrug transporter EmrE, an integral membrane protein

Manfred Schwaiger, Mario Lebendiker, Hagit Yerushalmi, Murray Coles, Adriane Gröger, Christian Schwarz, Shimon Schuldiner, Horst Kessler*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

89 Scopus citations

Abstract

EmrE is an Escherichia coli multidrug transport protein that confers resistance to a wide range of toxicants by active transport across the bacterial cell membrane. The highly hydrophobic polytopic integral membrane protein has been purified and studied in its full-length form by high- resolution NMR spectroscopy in a mixture of chloroform/methanol/water (6:6: 1, by vol.). Full activity is maintained after reconstitution of the protein into proteoliposomes from this solvent mixture. A series of heteronuclear (1H-15N) two- and three-dimensional experiments, as well as triple resonance experiments, were applied to the 110-residue protein and led to the assignment of the 1H, 15N and a large part of the 13C backbone resonances as well as many of the sidechain resonances. A preliminary analysis of the secondary structure, based on sequential NOE connectivities, deviation of chemical shifts from random coil values and 3J(NH-Hα) coupling constants supports a model where the protein forms four α-helices between residues 4-26 (TM1), 32-53 (TM2), 58-76 (TM3) and 85-106 (TM4). For the residues of helices TM2 and TM3 a significant line broadening occurs due to slow conformational processes.

Original languageEnglish
Pages (from-to)610-619
Number of pages10
JournalEuropean Journal of Biochemistry
Volume254
Issue number3
DOIs
StatePublished - 15 Jun 1998

Keywords

  • EmrE protein
  • Membrane protein
  • Multidrug resistance
  • NMR
  • Secondary structure

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