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Influence of detergents on the activity of the ABC transporter LmrA

Infed, N., Hanekop, N., Driessen, A. J. M., Smits, S. H. J. & Schmitt, L., Sep-2011, In : Biochimica et Biophysica Acta-Biomembranes. 1808, 9, p. 2313-2321 9 p.

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  • Influence of detergents on the activity of the ABC transporter LmrA

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DOI

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. (C) 2011 Elsevier B.V. All rights reserved.

Original languageEnglish
Pages (from-to)2313-2321
Number of pages9
JournalBiochimica et Biophysica Acta-Biomembranes
Volume1808
Issue number9
Publication statusPublished - Sep-2011

    Keywords

  • Detergent screen, ABC transporter, Functional reconstitution, ATPase activity, INTEGRAL MEMBRANE-PROTEINS, MULTIDRUG TRANSPORTER, P-GLYCOPROTEIN, LACTOCOCCUS-LACTIS, ATP HYDROLYSIS, BINDING, PURIFICATION, EXPRESSION, RESISTANCE, SOLUBILIZATION

ID: 5372361