FCS study of the thermodynamics of membrane protein insertion into the lipid bilayer chaperoned by fluorinated surfactants.

Biophysical Journal
Yevgen O PosokhovA S Ladokhin

Abstract

Experimental determination of the free energy (DeltaG) stabilizing the structure of membrane proteins (MPs) in their native environment has been hampered by the aggregation and precipitation of MPs outside the lipid bilayer. We recently demonstrated that the latter process can be prevented by the use of fluorinated surfactants, FTACs, that act as chaperones for MP insertion without partitioning in the membrane themselves. Here we combine the advantages of the chaperone-like ability of FTACs with the sensitivity of fluorescence correlation spectroscopy measurements to determine DeltaG of bilayer insertion of model MPs. First, we calibrate our approach by examining the effects of chaperoned insertion on DeltaG of transmembrane insertion of Annexin B12. We find that a shorter-chained surfactant, FTAC-C6, for which the working concentration range of 0.05-0.2 mM falls below CMC = 0.33 mM, has a mild effect on an apparent DeltaG. In contrast, additions of a longer-chained FTAC-C8 (CMC = 0.03 mM) result in a steep and nonlinear concentration dependence of DeltaG. We then apply the same methodology to the pH-triggered insertion of diphtheria toxin T-domain, which is known to be affected by nonproductive aggregation in solution. We find...Continue Reading

References

Oct 1, 1996·Nature Structural Biology·W C Wimley, S H White
Dec 24, 1996·Proceedings of the National Academy of Sciences of the United States of America·C TribetJ L Popot
Jul 4, 2001·The Journal of Biological Chemistry·S H WhiteK Hristova
May 12, 2004·Biochemistry·Alexey S Ladokhin, Stephen H White
Jun 9, 2004·Biochemistry·Alexey S LadokhinStephen H White
May 29, 2007·Journal of Molecular Biology·Mónica Fernández-VidalStephen H White

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Citations

Jul 27, 2010·Journal of Molecular Biology·Mykola V RodninAlexey S Ladokhin
Feb 5, 2015·The Journal of Membrane Biology·Jose C Flores-CanalesMaria Kurnikova
Oct 5, 2014·The Journal of Membrane Biology·Mauricio Vargas-UribeAlexey S Ladokhin
Jan 10, 2012·Journal of Molecular Biology·Alexander KyrychenkoAlexey S Ladokhin
Nov 3, 2016·Biophysical Journal·Mykola V RodninAlexey S Ladokhin
Mar 9, 2020·Analytical and Bioanalytical Chemistry·Xu FuChristopher I Richards
Oct 15, 2019·Frontiers in Molecular Biosciences·H RaghuramanAnindita Das
Feb 18, 2020·Methods : a Companion to Methods in Enzymology·Clarisse FaugierAnass Jawhari

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