Influence of detergents on the activity of the ABC transporter LmrA

Biochim Biophys Acta. 2011 Sep;1808(9):2313-21. doi: 10.1016/j.bbamem.2011.05.016. Epub 2011 May 30.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP-Binding Cassette Transporters / chemistry
  • Adenosine Triphosphatases / chemistry
  • Adenosine Triphosphate / chemistry
  • Bacterial Proteins / chemistry*
  • Chromatography / methods
  • Detergents / chemistry*
  • Drug Resistance, Multiple
  • Escherichia coli / metabolism
  • Hydrolysis
  • Kinetics
  • Lactococcus lactis / metabolism*
  • Liposomes / chemistry
  • Multidrug Resistance-Associated Proteins / chemistry*
  • Rhodamine 123 / pharmacology
  • Substrate Specificity
  • Temperature

Substances

  • ATP-Binding Cassette Transporters
  • Bacterial Proteins
  • Detergents
  • Liposomes
  • LmrA protein, Lactococcus lactis
  • Multidrug Resistance-Associated Proteins
  • Rhodamine 123
  • Adenosine Triphosphate
  • Adenosine Triphosphatases