FoldAffinity: binding affinities from nDSF experiments

Abstract Differential scanning fluorimetry (DSF) using the inherent fluorescence of proteins (nDSF) is a popular technique to evaluate thermal protein stability in different conditions (e.g. buffer, pH). In many cases, ligand binding increases thermal stability of a protein and often this can be det...

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Autores principales:	Stephan Niebling, Osvaldo Burastero, Jérôme Bürgi, Christian Günther, Lucas A. Defelipe, Simon Sander, Ellen Gattkowski, Raghavendra Anjanappa, Matthias Wilmanns, Sebastian Springer, Henning Tidow, María García-Alai
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e0885ebc8fc94564a375e92b59216b89
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spelling	oai:doaj.org-article:e0885ebc8fc94564a375e92b59216b892021-12-02T14:29:15ZFoldAffinity: binding affinities from nDSF experiments10.1038/s41598-021-88985-z2045-2322https://doaj.org/article/e0885ebc8fc94564a375e92b59216b892021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-88985-zhttps://doaj.org/toc/2045-2322Abstract Differential scanning fluorimetry (DSF) using the inherent fluorescence of proteins (nDSF) is a popular technique to evaluate thermal protein stability in different conditions (e.g. buffer, pH). In many cases, ligand binding increases thermal stability of a protein and often this can be detected as a clear shift in nDSF experiments. Here, we evaluate binding affinity quantification based on thermal shifts. We present four protein systems with different binding affinity ligands, ranging from nM to high μM. Our study suggests that binding affinities determined by isothermal analysis are in better agreement with those from established biophysical techniques (ITC and MST) compared to apparent K d s obtained from melting temperatures. In addition, we describe a method to optionally fit the heat capacity change upon unfolding ( $$\Delta {C}_{p}$$ Δ C p ) during the isothermal analysis. This publication includes the release of a web server for easy and accessible application of isothermal analysis to nDSF data.Stephan NieblingOsvaldo BurasteroJérôme BürgiChristian GüntherLucas A. DefelipeSimon SanderEllen GattkowskiRaghavendra AnjanappaMatthias WilmannsSebastian SpringerHenning TidowMaría García-AlaiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-17 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Stephan Niebling Osvaldo Burastero Jérôme Bürgi Christian Günther Lucas A. Defelipe Simon Sander Ellen Gattkowski Raghavendra Anjanappa Matthias Wilmanns Sebastian Springer Henning Tidow María García-Alai FoldAffinity: binding affinities from nDSF experiments
description	Abstract Differential scanning fluorimetry (DSF) using the inherent fluorescence of proteins (nDSF) is a popular technique to evaluate thermal protein stability in different conditions (e.g. buffer, pH). In many cases, ligand binding increases thermal stability of a protein and often this can be detected as a clear shift in nDSF experiments. Here, we evaluate binding affinity quantification based on thermal shifts. We present four protein systems with different binding affinity ligands, ranging from nM to high μM. Our study suggests that binding affinities determined by isothermal analysis are in better agreement with those from established biophysical techniques (ITC and MST) compared to apparent K d s obtained from melting temperatures. In addition, we describe a method to optionally fit the heat capacity change upon unfolding ( $$\Delta {C}_{p}$$ Δ C p ) during the isothermal analysis. This publication includes the release of a web server for easy and accessible application of isothermal analysis to nDSF data.
format	article
author	Stephan Niebling Osvaldo Burastero Jérôme Bürgi Christian Günther Lucas A. Defelipe Simon Sander Ellen Gattkowski Raghavendra Anjanappa Matthias Wilmanns Sebastian Springer Henning Tidow María García-Alai
author_facet	Stephan Niebling Osvaldo Burastero Jérôme Bürgi Christian Günther Lucas A. Defelipe Simon Sander Ellen Gattkowski Raghavendra Anjanappa Matthias Wilmanns Sebastian Springer Henning Tidow María García-Alai
author_sort	Stephan Niebling
title	FoldAffinity: binding affinities from nDSF experiments
title_short	FoldAffinity: binding affinities from nDSF experiments
title_full	FoldAffinity: binding affinities from nDSF experiments
title_fullStr	FoldAffinity: binding affinities from nDSF experiments
title_full_unstemmed	FoldAffinity: binding affinities from nDSF experiments
title_sort	foldaffinity: binding affinities from ndsf experiments
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/e0885ebc8fc94564a375e92b59216b89
work_keys_str_mv	AT stephanniebling foldaffinitybindingaffinitiesfromndsfexperiments AT osvaldoburastero foldaffinitybindingaffinitiesfromndsfexperiments AT jeromeburgi foldaffinitybindingaffinitiesfromndsfexperiments AT christiangunther foldaffinitybindingaffinitiesfromndsfexperiments AT lucasadefelipe foldaffinitybindingaffinitiesfromndsfexperiments AT simonsander foldaffinitybindingaffinitiesfromndsfexperiments AT ellengattkowski foldaffinitybindingaffinitiesfromndsfexperiments AT raghavendraanjanappa foldaffinitybindingaffinitiesfromndsfexperiments AT matthiaswilmanns foldaffinitybindingaffinitiesfromndsfexperiments AT sebastianspringer foldaffinitybindingaffinitiesfromndsfexperiments AT henningtidow foldaffinitybindingaffinitiesfromndsfexperiments AT mariagarciaalai foldaffinitybindingaffinitiesfromndsfexperiments
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FoldAffinity: binding affinities from nDSF experiments

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