title: Influence of detergents on the activity of the ABC transporter LmrA.
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.
authors.affiliation: Institute of Biochemistry, Universitaetstr 1; Heinrich-Heine-University, 40225 Duesseldorf, Germany.
authors.forename: Nacera
authors.initials: N
authors.lastname: Infed
authors.forename: Nils
authors.initials: N
authors.lastname: Hanekop
authors.forename: Arnold J M
authors.initials: AJ
authors.lastname: Driessen
authors.forename: Sander H J
authors.initials: SH
authors.lastname: Smits
authors.forename: Lutz
authors.initials: L
authors.lastname: Schmitt
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000737:chemistry; D000251:Adenosine Triphosphatases / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry*; D002845:Chromatography / Q000379:methods; D003902:Detergents / Q000737:chemistry*; D018432:Drug Resistance, Multiple; D004926:Escherichia coli / Q000378:metabolism; D006868:Hydrolysis; D007700:Kinetics; D013294:Lactococcus lactis / Q000378:metabolism*; D008081:Liposomes / Q000737:chemistry; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000737:chemistry*; D020112:Rhodamine 123 / Q000494:pharmacology; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D018528:ATP-Binding Cassette Transporters; D000251:Adenosine Triphosphatases; D000255:Adenosine Triphosphate; D001426:Bacterial Proteins; D003902:Detergents; D008081:Liposomes; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D020112:Rhodamine 123; C101824:LmrA protein, Lactococcus lactis