EmrE dimerization depends on membrane environment |
| |
Authors: | Supratik Dutta Emma A Morrison Katherine A Henzler-Wildman |
| |
Institution: | Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, 660 S. Euclid Ave., Box 8231, St. Louis, MO 63110, USA |
| |
Abstract: | The small multi-drug resistant (SMR) transporter EmrE functions as a homodimer. Although the small size of EmrE would seem to make it an ideal model system, it can also make it challenging to work with. As a result, a great deal of controversy has surrounded even such basic questions as the oligomeric state. Here we show that the purified protein is a homodimer in isotropic bicelles with a monomer–dimer equilibrium constant (KMD2D) of 0.002–0.009 mol% for both the substrate-free and substrate-bound states. Thus, the dimer is stabilized in bicelles relative to detergent micelles where the KMD2D is only 0.8–0.95 mol% (Butler et al. 2004). In dilauroylphosphatidylcholine (DLPC) liposomes KMD2D is 0.0005–0.0008 mol% based on Förster resonance energy transfer (FRET) measurements, slightly tighter than bicelles. These results emphasize the importance of the lipid membrane in influencing dimer affinity. |
| |
Keywords: | SMR small multidrug resistance EmrE Escherichia coli SMR protein FRET Fö rster resonance energy transfer AUC analytical ultra centrifugation DDM dodecylmaltoside DHPC 1 2-dihexanoyl-sn-glycero-3-phosphocholine DLPC 1 2-dilauroyl-sn-glycero-3-phosphocholine TPP+ tetraphenylphosphonium + TM transmembrane |
本文献已被 ScienceDirect 等数据库收录! |