Probing the molecular dynamics of the ABC multidrug transporter LmrA by deuterium solid-state nuclear magnetic resonance

Biochemistry. 2007 Mar 20;46(11):3075-83. doi: 10.1021/bi062109a. Epub 2007 Feb 16.

Abstract

The molecular dynamics of the 64 kDa ABC multidrug efflux pump LmrA from Lactococcus lactis within lipid membranes has been investigated by deuterium solid-state NMR. Deuteriomethyl-labeled alanine has been used to probe global protein backbone dynamics. A comparison of static deuterium NMR spectra of full-length LmrA in the resting state and its isolated transmembrane domain revealed a high mobility for the nucleotide binding domains. Their motional freedom is restricted upon ATP binding as seen for LmrA in complex with AMP-PNP, a nonhydrolyzable ATP analogue. LmrA returns to full motional flexibility in the posthydrolysis, vanadate-trapped state. These experiments provide insight into the molecular dynamics of a full-length ABC transporter during the catalytic cycle. Data are discussed in the context of known biochemical data and structural models of ABC transporters.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alanine / chemistry
  • Amino Acid Sequence
  • Bacterial Proteins / chemistry*
  • Deuterium
  • Electrophoresis, Polyacrylamide Gel
  • Molecular Sequence Data
  • Multidrug Resistance-Associated Proteins / chemistry*
  • Nuclear Magnetic Resonance, Biomolecular
  • Phosphorus Isotopes

Substances

  • Bacterial Proteins
  • LmrA protein, Lactococcus lactis
  • Multidrug Resistance-Associated Proteins
  • Phosphorus Isotopes
  • Deuterium
  • Alanine