Probing the molecular dynamics of the ABC multidrug transporter LmrA by deuterium solid-state nuclear magnetic resonance.
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.
Siarheyeva
Alena
A
Institute for Biophysical Chemistry and Centre for Biomolecular Magnetic Resonance, J. W. Goethe Universität, Frankfurt am Main, Germany.
Lopez
Jakob J
JJ
Lehner
Ines
I
Hellmich
Ute A
UA
van Veen
Hendrik W
HW
Glaubitz
Clemens
C
D000409:Alanine / Q000737:chemistry; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry*; D003903:Deuterium; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular; D010759:Phosphorus Isotopes
D000409:Alanine; D001426:Bacterial Proteins; D003903:Deuterium; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D010759:Phosphorus Isotopes; C101824:LmrA protein, Lactococcus lactis