Influence of detergents on the activity of the ABC transporter LmrA |
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Authors: | Infed Nacera Hanekop Nils Driessen Arnold J M Smits Sander H J Schmitt Lutz |
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Institution: | Institute of Biochemistry, Universitaetstr 1; Heinrich-Heine-University, 40225 Duesseldorf, Germany. |
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Abstract: | The ABC transporter LmrA from Lactococcus lactis has been intensively studied and a role in multidrug resistance was proposed. Here, we performed a comprehensive detergent screen to analyze the impact of detergents for a successful solubilization, purification and retention of functional properties of this ABC transporter. Our screen revealed the preference of LmrA for zwitterionic detergents. In detergent solution, LmrA purified with FC-16 was highly active with respect to ATPase activity, which could be stimulated by a substrate (rhodamine 123) of LmrA. Both, high ATPase activity and substrate stimulation were not detected for LmrA solubilized in DDM. Interestingly, reconstituted LmrA showed an opposite behavior, with a high basal ATPase activity and stimulation by rhodamine 123 for a DDM-reconstituted, but only low ATPase activity and no substrate stimulation for a FC-16 reconstituted sample. |
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