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Published on: November 29, 2014
Amino acid type selective isotope labelling of the multidrug ABC transporter LmrA for solid-state NMR studies
A James Mason1, Alena Siarheyeva, Winfried Haase
1Centre for Biomolecular Magnetic Resonance and Institut für Biophysikalische Chemie, J.W. Goethe Universität, Marie-Curie Str. 9, D-60439 Frankfurt am Main, Germany.
Abstract:
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.
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