The reconstitution and activity of the small multidrug transporter EmrE is modulated by non-bilayer lipid composition

Journal of Molecular Biology
Paul CurnowPaula J Booth

Abstract

The ability of multidrug transport proteins within biological membranes to recognise a diverse array of substrates is a fundamental aspect of antibiotic resistance. Detailed information on the mechanisms of recognition and transport can be provided only by in vitro studies in reconstituted bilayer systems. We describe the controlled, efficient reconstitution of the small multidrug transporter EmrE in a simple model membrane and investigate the effect of non-bilayer lipids on this process. Transport activity is impaired, in line with an increase in the lateral pressure within the bilayer. We demonstrate the potential of this lateral pressure modulation method as a general approach to the folding and assembly of membrane proteins in vitro, by recovering functional transporter from a partly denatured state. Our results highlight the importance of optimising reconstitution procedures and bilayer lipid composition in studies of membrane transporters. This is particularly pertinent for multidrug proteins, and we show that the use of a sub-optimal lipid bilayer environment or reconstitution method could lead to incorrect information on protein activity.

References

Mar 1, 1990·FEMS Microbiology Letters·A S PurewalM Midgley
Jun 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·S M Gruner
Aug 4, 1998·European Journal of Biochemistry·M SchwaigerH Kessler
Nov 18, 1998·Biochemistry·J KnolB Poolman
Sep 2, 1999·Trends in Biochemical Sciences·C M Dobson
Jul 26, 2000·Proceedings of the National Academy of Sciences of the United States of America·G S AttardS Jackowski
Nov 25, 2000·Biochimica Et Biophysica Acta·M le MaireJ V Moller
Mar 30, 2001·The Journal of Biological Chemistry·H YerushalmiS Schuldiner
Dec 12, 2001·Science·M A SchumacherR G Brennan
Jan 19, 2002·Critical Reviews in Biochemistry and Molecular Biology·P J BoothM Lorch
May 16, 2002·Biochemistry·Ana Vitória BotelhoMichael F Brown
Jun 11, 2002·Journal of Molecular Biology·Wim Meijberg, Paula J Booth
Sep 11, 2002·Proceedings of the National Academy of Sciences of the United States of America·Misha SoskineShimon Schuldiner
Nov 8, 2002·The Journal of Biological Chemistry·Tip W LooDavid M Clarke
Mar 26, 2003·Biochemistry·Ellen M Howard, Paul D Roepe
May 27, 2003·The Journal of Biological Chemistry·Jason S PatzlaffBert Poolman
Jun 5, 2003·Nature Reviews. Molecular Cell Biology·Valerie Daggett, Alan Fersht
Aug 30, 2003·Journal of Molecular Biology·Christopher G TateJoyce M Baldwin
Nov 25, 2003·The EMBO Journal·Iban Ubarretxena-BelandiaChristopher G Tate
Feb 3, 2004·Proceedings of the National Academy of Sciences of the United States of America·Yael ElbazShimon Schuldiner
Feb 19, 2004·Proceedings of the National Academy of Sciences of the United States of America·Che Ma, Geoffrey Chang
Sep 8, 2004·Journal of Molecular Biology·Samantha J AllenPaula J Booth

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Citations

Mar 1, 2006·Naunyn-Schmiedeberg's Archives of Pharmacology·Daniel BastingClemens Glaubitz
Jun 21, 2011·Journal of the American Chemical Society·Heedeok Hong, James U Bowie
Oct 12, 2007·The Journal of Biological Chemistry·Aixin YanChristian R H Raetz
Nov 26, 2010·Protein Engineering, Design & Selection : PEDS·Dilip K DebnathDaniel E Otzen
Dec 7, 2010·Journal of the Royal Society, Interface·Mario OrsiJonathan W Essex
Nov 27, 2008·BMC Biochemistry·Kalypso CharalambousPaula J Booth
Oct 14, 2011·The Journal of Chemical Physics·J J López CascalesR D Porasso
Feb 7, 2013·Chemistry and Physics of Lipids·Safia NathooElmar J Prenner
Nov 22, 2011·Current Opinion in Structural Biology·Katherine Henzler-Wildman
Jan 28, 2009·Biochemical and Biophysical Research Communications·Mikio TanabeBernadette Byrne
Aug 12, 2015·Chemistry and Physics of Lipids·Pouyan Khakbaz, Jeffery B Klauda
Sep 8, 2004·Journal of Molecular Biology·Samantha J AllenPaula J Booth
Apr 1, 2014·Biochimica Et Biophysica Acta·Supratik DuttaKatherine A Henzler-Wildman
Oct 30, 2012·Biochimica Et Biophysica Acta·Karsten MörsClemens Glaubitz
Aug 25, 2009·Journal of Molecular Biology·David MillerPaula J Booth
Nov 22, 2011·Biochimica Et Biophysica Acta·Nicola J Harris, Paula J Booth
Oct 23, 2018·Soft Matter·Michail PalaiokostasMario Orsi
May 24, 2017·Annual Review of Biophysics·Michael F Brown
Oct 17, 2014·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·C N C CostaE M S Castanheira
Oct 16, 2020·Protein Science : a Publication of the Protein Society·Karolina Corin, James U Bowie
Feb 8, 2020·Biophysical Journal·Brian P JoseyMathias Lösche
Nov 21, 2012·Biochemistry·Michael F Brown
Dec 19, 2007·The Journal of Physical Chemistry. B·Mario OrsiJonathan W Essex
May 11, 2006·Chemical Reviews·Kevin R Mackenzie

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