SDS-induced multi-stage unfolding of a small globular protein through different denatured states revealed by single-molecule fluorescence.

Chemical Science
Georg KrainerDaniel Otzen

Abstract

Ionic surfactants such as sodium dodecyl sulfate (SDS) unfold proteins in a much more diverse yet effective way than chemical denaturants such as guanidium chloride (GdmCl). But how these unfolding processes compare on a molecular level is poorly understood. Here, we address this question by scrutinising the unfolding pathway of the globular protein S6 in SDS and GdmCl with single-molecule Förster resonance energy transfer (smFRET) spectroscopy. We show that the unfolding mechanism in SDS is strikingly different and convoluted in comparison to denaturation in GdmCl. In contrast to the reversible two-state unfolding behaviour in GdmCl characterised by kinetics on the timescale of seconds, SDS demonstrated not one, but four distinct regimes of interactions with S6, dependent on the surfactant concentration. At ≤1 mM SDS, S6 and surfactant molecules form quasi-micelles on a minute timescale; at millimolar [SDS], the protein denatures through an unfolded/denatured ensemble of highly heterogeneous states on a multi-second timescale; at tens of millimolar of SDS, the protein unfolds into a micelle-packed conformation on the second timescale; and >50 mM SDS, the protein unfolds with millisecond timescale dynamics. We propose a detaile...Continue Reading

References

Aug 1, 1997·Trends in Biochemical Sciences·J ClarkeA R Fersht
Oct 16, 1999·Proceedings of the National Academy of Sciences of the United States of America·D E Otzen, M Oliveberg
Apr 28, 2001·Biophysical Journal·J T Vivian, P R Callis
Oct 26, 2001·Journal of Molecular Biology·D E Otzen, M Oliveberg
Feb 6, 2002·Journal of Molecular Biology·Daniel E Otzen, Mikael Oliveberg
Sep 27, 2002·Current Opinion in Biotechnology·Ole KirkClaus Crone Fuglsang
Jun 16, 2005·Biochimica Et Biophysica Acta·Daniel E Otzen
Dec 18, 2007·Biochimica Et Biophysica Acta·Daniel E OtzenPankaj Sehgal
Dec 18, 2007·Journal of Molecular Biology·Catherine MichauxGilbert G Privé
Oct 3, 2008·Journal of Colloid and Interface Science·Janaky NarayananJayesh R Bellare
Jun 16, 2009·Journal of Molecular Biology·Kell K AndersenDaniel Otzen
Jun 15, 2010·Journal of Molecular Biology·Lise GiehmDaniel E Otzen
Jan 12, 2011·Annual Review of Physical Chemistry·Jeremy L England, Gilad Haran
Mar 15, 2011·Biochimica Et Biophysica Acta·Daniel Otzen
Mar 3, 2012·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Volodymyr KudryavtsevDon C Lamb
Jan 10, 2013·Annual Review of Physical Chemistry·Deepak R Canchi, Angel E García
Jan 15, 2013·Current Opinion in Structural Biology·Benjamin Schuler, Hagen Hofmann
Aug 27, 2015·Soft Matter·Yang SunCristiano L P Oliveira
Sep 2, 2016·Journal of the American Chemical Society·Alessandro BorgiaBenjamin Schuler
Apr 27, 2017·Biophysical Journal·Jørn Døvling KaspersenJan Skov Pedersen
Nov 13, 2017·Journal of Molecular Biology·Georg KrainerMichael Schlierf
Dec 21, 2017·Biophysical Journal·Henriette Gavlshøj MortensenJan Skov Pedersen
Feb 22, 2018·Nature·Alessandro BorgiaBenjamin Schuler
Jul 20, 2019·Current Opinion in Structural Biology·Georg KrainerMichael Schlierf
Mar 8, 2020·Nature Communications·Hugo SanabriaClaus A M Seidel
Nov 22, 2019·Chemical Science·Jannik Nedergaard PedersenJan Skov Pedersen

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Methods Mentioned

BETA
circular dichroism
FRET
isothermal titration calorimetry
size-exclusion chromatography
fluorescence spectroscopy

Software Mentioned

Matlab scripts
Mathworks
GenScript
smFRET

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