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Daniel Otzen

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

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SDS-induced multi-stage unfolding of a small globular protein through different denatured states revealed by single-molecule fluorescence. / Krainer, Georg; Hartmann, Andreas; Bogatyr, Vadim et al.

In: Chemical Science, Vol. 11, No. 34, 09.2020, p. 9141-9153.

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Krainer G, Hartmann A, Bogatyr V, Nielsen J, Schlierf M, Otzen DE. SDS-induced multi-stage unfolding of a small globular protein through different denatured states revealed by single-molecule fluorescence. Chemical Science. 2020 Sep;11(34):9141-9153. doi: 10.1039/d0sc02100h

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Krainer, Georg ; Hartmann, Andreas ; Bogatyr, Vadim et al. / SDS-induced multi-stage unfolding of a small globular protein through different denatured states revealed by single-molecule fluorescence. In: Chemical Science. 2020 ; Vol. 11, No. 34. pp. 9141-9153.

Bibtex

@article{c56ea3adfd5649d38ab69e9df93705b8,
title = "SDS-induced multi-stage unfolding of a small globular protein through different denatured states revealed by single-molecule fluorescence",
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 F{\"o}rster 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 detailed model for multi-stage unfolding of S6 in SDS, which involves at least three different types of denatured states with different level of compactness and dynamics and a continually changing landscape of interactions between protein and surfactant. Our results highlight the great potential of single-molecule fluorescence as a direct probe of nanoscale protein structure and dynamics in chemically complex surfactant environments.",
author = "Georg Krainer and Andreas Hartmann and Vadim Bogatyr and Janni Nielsen and Michael Schlierf and Otzen, {Daniel E.}",
year = "2020",
month = sep,
doi = "10.1039/d0sc02100h",
language = "English",
volume = "11",
pages = "9141--9153",
journal = "Chemical Science",
issn = "2041-6520",
publisher = "royal society of chemistry",
number = "34",

}

RIS

TY - JOUR

T1 - SDS-induced multi-stage unfolding of a small globular protein through different denatured states revealed by single-molecule fluorescence

AU - Krainer, Georg

AU - Hartmann, Andreas

AU - Bogatyr, Vadim

AU - Nielsen, Janni

AU - Schlierf, Michael

AU - Otzen, Daniel E.

PY - 2020/9

Y1 - 2020/9

N2 - 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 detailed model for multi-stage unfolding of S6 in SDS, which involves at least three different types of denatured states with different level of compactness and dynamics and a continually changing landscape of interactions between protein and surfactant. Our results highlight the great potential of single-molecule fluorescence as a direct probe of nanoscale protein structure and dynamics in chemically complex surfactant environments.

AB - 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 detailed model for multi-stage unfolding of S6 in SDS, which involves at least three different types of denatured states with different level of compactness and dynamics and a continually changing landscape of interactions between protein and surfactant. Our results highlight the great potential of single-molecule fluorescence as a direct probe of nanoscale protein structure and dynamics in chemically complex surfactant environments.

UR - http://www.scopus.com/inward/record.url?scp=85090763998&partnerID=8YFLogxK

U2 - 10.1039/d0sc02100h

DO - 10.1039/d0sc02100h

M3 - Journal article

C2 - 34123163

AN - SCOPUS:85090763998

VL - 11

SP - 9141

EP - 9153

JO - Chemical Science

JF - Chemical Science

SN - 2041-6520

IS - 34

ER -