MAS solid-state NMR studies on the multidrug transporter EmrE.
We study the uniformly 13C,15N isotopically enriched Escherichia coli multidrug resistance transporter EmrE using MAS solid-state NMR. Solid-state NMR can provide complementary structural information as the method allows studying membrane proteins in their native environment as no detergent is required for reconstitution. We compare the spectra obtained from wildtype EmrE to those obtained from the mutant EmrE-E14C. To resolve the critical amino acid E14, glutamic/aspartic acid selective experiments are carried out. These experiments allow to assign the chemical shift of the carboxylic carbon of E14. In addition, spectra are analyzed which are obtained in the presence and absence of the ligand TPP+.
Agarwal
Vipin
V
Leibniz-Institut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, D-13125 Berlin, Germany.
Fink
Uwe
U
Schuldiner
Shimon
S
Reif
Bernd
B
D017920:Antiporters / Q000737:chemistry* / Q000648:ultrastructure; D002247:Carbon Isotopes; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000648:ultrastructure; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008854:Microscopy, Electron; D009154:Mutation; D009587:Nitrogen Isotopes; D009861:Onium Compounds / Q000378:metabolism; D009943:Organophosphorus Compounds / Q000378:metabolism; D011485:Protein Binding
D017920:Antiporters; D002247:Carbon Isotopes; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D009587:Nitrogen Isotopes; D009861:Onium Compounds; D009943:Organophosphorus Compounds; C075436:EmrE protein, E coli; C013289:tetraphenylphosphonium