Please use this identifier to cite or link to this item:
http://dx.doi.org/10.25673/85885
Title: | Monitoring protein unfolding transitions by NMR-spectroscopy |
Author(s): | Dreydoppel, Matthias Balbach, Jochen Weininger, Ulrich |
Issue Date: | 2022 |
Type: | Article |
Language: | English |
Abstract: | NMR-spectroscopy has certain unique advantages for recording unfolding transitions of proteins compared e.g. to optical methods. It enables per-residue monitoring and separate detection of the folded and unfolded state as well as possible equilibrium intermediates. This allows a detailed view on the state and cooperativity of folding of the protein of interest and the correct interpretation of subsequent experiments. Here we summarize in detail practical and theoretical aspects of such experiments. Certain pitfalls can be avoided, and meaningful simplification can be made during the analysis. Especially a good understanding of the NMR exchange regime and relaxation properties of the system of interest is beneficial. We show by a global analysis of signals of the folded and unfolded state of GB1 how accurate values of unfolding can be extracted and what limits different NMR detection and unfolding methods. E.g. commonly used exchangeable amides can lead to a systematic under determination of the thermodynamic protein stability. We give several perspectives of how to deal with more complex proteins and how the knowledge about protein stability at residue resolution helps to understand protein properties under crowding conditions, during phase separation and under high pressure. |
URI: | https://opendata.uni-halle.de//handle/1981185920/87838 http://dx.doi.org/10.25673/85885 |
Open Access: | Open access publication |
License: | (CC BY 4.0) Creative Commons Attribution 4.0 |
Sponsor/Funder: | Publikationsfonds MLU |
Journal Title: | Journal of biomolecular NMR |
Publisher: | Springer Science + Business Media B.V |
Publisher Place: | Dordrecht [u.a.] |
Volume: | 76 |
Original Publication: | 10.1007/s10858-021-00389-3 |
Page Start: | 3 |
Page End: | 15 |
Appears in Collections: | Open Access Publikationen der MLU |
Files in This Item:
File | Description | Size | Format | |
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Dreydoppel2022_Article_MonitoringProteinUnfoldingTran.pdf | 2.06 MB | Adobe PDF | View/Open |