Browsing by Author "Chew, Raymond"
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- ItemOpen AccessComparisons of EmrE Purification Methods(2013-04-30) Chew, Raymond; Turner, RaymondDue to the difficulties of membrane protein expression, isolation, and solubilisation, purification yields are typically lower than most soluble proteins. A common tactic to increase yield is the use of affinity tags for co-purification of the attached protein. EmrE, a small bacterial multidrug efflux protein was examined to characterize changes in structure and function due to a His6 affinity tag. Fluorescence spectroscopy revealed the presence of a relatively hydrophobic fold in tagged EmrE as well as slower solvent dynamics within the folded protein. In regards to functionality, the untagged EmrE displayed higher ligand binding affinities to ethidium and methyl viologen in vitro and also higher transport activity in vivo.