Folding kinetics of an alpha helical membrane protein in phospholipid bilayer vesicles

Journal of Molecular Biology
Samantha J AllenP J Booth

Abstract

We report a detailed kinetic study of the folding of an alpha-helical membrane protein in a lipid bilayer environment. SDS denatured bacteriorhodopsin was folded directly into phosphatidylcholine lipid vesicles by stopped-flow mixing. The folding kinetics were monitored with millisecond time resolution by time-resolving changes in protein fluorescence as well as in the absorption of the retinal chromophore. The kinetics were similar to those previously reported for folding bacteriorhodopsin in detergent or lipid micelles, except for the presence of an additional apoprotein intermediate. We suggest this intermediate is a result of the greater internal two-dimensional pressure present in these lipid vesicles as compared to micelles. These results lay the groundwork for future studies aimed at understanding the mechanistic origin of the effect of lipid bilayer properties on protein folding. Furthermore, the use of biologically relevant phosphatidylcholine lipids, together with a straightforward rapid mixing process to initiate the folding reaction, means the method is generally applicable, and thus paves the way for an improved understanding of the in vitro folding of transmembrane alpha-helical proteins.

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Citations

Jan 17, 2012·Biochemistry·Venkatramanan KrishnamaniJanos K Lanyi
Jun 25, 2008·Annual Review of Biophysics·Stephen H White, Gunnar von Heijne
Oct 4, 2005·Protein Science : a Publication of the Protein Society·Jesper E MogensenDaniel E Otzen
Nov 17, 2004·Journal of Molecular Biology·Mark Lorch, Paula J Booth
Sep 8, 2004·Journal of Molecular Biology·Samantha J AllenPaula J Booth
Jul 4, 2006·Current Opinion in Structural Biology·Paula J Booth, Paul Curnow
Jan 24, 2006·Journal of Molecular Biology·Emma L R ComptonPaula J Booth
Mar 16, 2011·Biophysical Journal·Venkatramanan Krishnamani, Janos K Lanyi
Aug 25, 2009·Journal of Molecular Biology·David MillerPaula J Booth
Nov 26, 2014·Quarterly Reviews of Biophysics·Jonathan P Schlebach, Charles R Sanders
Dec 18, 2002·Journal of the American Society for Mass Spectrometry·Raino K HansenIsaiah T Arkin
Oct 7, 2005·Chemistry and Physics of Lipids·Mandeep Singh BakshiGurinder Kaur
Sep 8, 2018·Biochemical Society Transactions·Nicola J HarrisPaula J Booth
Aug 16, 2017·Scientific Reports·Nicola J HarrisPaula J Booth
Nov 11, 2016·Nanotechnology·David M MillerPaula J Booth
Oct 16, 2020·Protein Science : a Publication of the Protein Society·Karolina Corin, James U Bowie
Jun 3, 2009·The Journal of Physical Chemistry. B·Jing ZhangC Ted Lee
May 30, 2013·The Journal of Physical Chemistry. B·Julia MuenznerEkaterina V Pletneva
Nov 1, 2007·Archives of Biochemistry and Biophysics·Ann Marie Stanley, Karen G Fleming
Jun 21, 2008·Journal of Molecular Biology·Annela M SeddonPaula J Booth
May 11, 2006·Chemical Reviews·Kevin R Mackenzie

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