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Quantifying the thermodynamics of protein unfolding using 2D NMR spectroscopy

A topic that has attracted considerable interest in recent years is the possibility to perform thermodynamic studies of proteins directly in-cell or in complex environments which mimic the cellular interior. Nuclear magnetic resonance (NMR) could be an attractive technique for these studies but its...

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Autores principales: Puglisi, Rita, Brylski, Oliver, Alfano, Caterina, Martin, Stephen R., Pastore, Annalisa, Temussi, Piero A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7116895/
https://www.ncbi.nlm.nih.gov/pubmed/33718626
http://dx.doi.org/10.1038/s42004-020-00358-1
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author Puglisi, Rita
Brylski, Oliver
Alfano, Caterina
Martin, Stephen R.
Pastore, Annalisa
Temussi, Piero A.
author_facet Puglisi, Rita
Brylski, Oliver
Alfano, Caterina
Martin, Stephen R.
Pastore, Annalisa
Temussi, Piero A.
author_sort Puglisi, Rita
collection PubMed
description A topic that has attracted considerable interest in recent years is the possibility to perform thermodynamic studies of proteins directly in-cell or in complex environments which mimic the cellular interior. Nuclear magnetic resonance (NMR) could be an attractive technique for these studies but its applicability has so far been limited by technical issues. Here, we demonstrate that 2D NMR methods can be successfully applied to measure thermodynamic parameters provided that a suitable choice of the residues used for the calculation is made. We propose a new parameter, named RAD, which reflects the level of protection of a specific amide proton in the protein core and can guide through the selection of the resonances. We also suggest a way to calibrate the volumes to become independent of technical limitations. The methodology we propose leads to stability curves comparable to that calculated from CD data and provides a new tool for thermodynamic measurements in complex environments.
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spelling pubmed-71168952021-03-13 Quantifying the thermodynamics of protein unfolding using 2D NMR spectroscopy Puglisi, Rita Brylski, Oliver Alfano, Caterina Martin, Stephen R. Pastore, Annalisa Temussi, Piero A. Commun Chem Article A topic that has attracted considerable interest in recent years is the possibility to perform thermodynamic studies of proteins directly in-cell or in complex environments which mimic the cellular interior. Nuclear magnetic resonance (NMR) could be an attractive technique for these studies but its applicability has so far been limited by technical issues. Here, we demonstrate that 2D NMR methods can be successfully applied to measure thermodynamic parameters provided that a suitable choice of the residues used for the calculation is made. We propose a new parameter, named RAD, which reflects the level of protection of a specific amide proton in the protein core and can guide through the selection of the resonances. We also suggest a way to calibrate the volumes to become independent of technical limitations. The methodology we propose leads to stability curves comparable to that calculated from CD data and provides a new tool for thermodynamic measurements in complex environments. Nature Publishing Group UK 2020-08-07 /pmc/articles/PMC7116895/ /pubmed/33718626 http://dx.doi.org/10.1038/s42004-020-00358-1 Text en © The Author(s) 2020 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Puglisi, Rita
Brylski, Oliver
Alfano, Caterina
Martin, Stephen R.
Pastore, Annalisa
Temussi, Piero A.
Quantifying the thermodynamics of protein unfolding using 2D NMR spectroscopy
title Quantifying the thermodynamics of protein unfolding using 2D NMR spectroscopy
title_full Quantifying the thermodynamics of protein unfolding using 2D NMR spectroscopy
title_fullStr Quantifying the thermodynamics of protein unfolding using 2D NMR spectroscopy
title_full_unstemmed Quantifying the thermodynamics of protein unfolding using 2D NMR spectroscopy
title_short Quantifying the thermodynamics of protein unfolding using 2D NMR spectroscopy
title_sort quantifying the thermodynamics of protein unfolding using 2d nmr spectroscopy
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7116895/
https://www.ncbi.nlm.nih.gov/pubmed/33718626
http://dx.doi.org/10.1038/s42004-020-00358-1
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