Amino acid type selective isotope labelling of the multidrug ABC transporter LmrA for solid-state NMR studies.
The ABC transporter LmrA in Lactococcus lactis confers resistance to a wide range of antibiotics and cytotoxic drugs and is a functional homologue of P-glycoprotein. Recently, solid-state NMR methods have shown potential for structural- and non-perturbing, site directed functional studies. These experiments require isotopic labelling of selected sites. We have developed a strategy to produce large quantities of selectively labelled LmrA reconstituted at a high density in lipid membranes. This makes the 64 kDa integral membrane protein LmrA and therefore the ABC transporter superfamily accessible to NMR analysis.
Mason
A James
AJ
Centre for Biomolecular Magnetic Resonance and Institut für Biophysikalische Chemie, J.W. Goethe Universität, Marie-Curie Str. 9, D-60439 Frankfurt am Main, Germany.
Siarheyeva
Alena
A
Haase
Winfried
W
Lorch
Mark
M
van Veen
Hendrik
H
Glaubitz
Clemens
C
D000596:Amino Acids / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000648:ultrastructure; D008854:Microscopy, Electron; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000737:chemistry* / Q000648:ultrastructure; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*
D000596:Amino Acids; D001426:Bacterial Proteins; D027425:ATP-Binding Cassette, Sub-Family C Proteins; C101824:LmrA protein, Lactococcus lactis