the Creative Commons Attribution 4.0 License.
the Creative Commons Attribution 4.0 License.
Anatomy of unfolding: The site-specific fold stability of Yfh1 measured by 2D NMR
Abstract. Most techniques allow detection of protein unfolding either by following the behaviour of single reporters or as an averaged all-or-none process. We recently added 2D NMR spectroscopy to the well-established techniques able to obtain information on the process of unfolding using resonances of residues in the hydrophobic core of a protein. Here, we questioned whether a detailed analysis of the individual stability curves from each resonance could provide additional site-specific information. We used the Yfh1 protein that has the unique feature to undergo both cold and heat denaturation at temperatures above water freezing at low ionic strength. We show that stability curves inconsistent with the average NMR curve from hydrophobic core residues mainly comprise exposed outliers that do nevertheless provide precious information. By monitoring both cold and heat denaturation of individual residues we gain knowledge on the process of cold denaturation and convincingly demonstrate that the two unfolding processes are intrinsically different.
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Preprint
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This preprint has been withdrawn.
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Interactive discussion
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SC1: 'Standards of MR', Gottfried Otting, 24 Oct 2020
- AC1: 'Otting observations', Piero Andrea Temussi, 29 Oct 2020
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RC1: 'mr-2020-24', Anonymous Referee #1, 24 Oct 2020
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AC2: 'reply to Anonymous observations', Piero Andrea Temussi, 29 Oct 2020
- AC3: 'revised answer to anonymous referee', Piero Andrea Temussi, 18 Dec 2020
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AC2: 'reply to Anonymous observations', Piero Andrea Temussi, 29 Oct 2020
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RC2: 'mr-2020-24', Dmitry Korzhnev, 27 Oct 2020
- AC4: 'answer to reviewer 2', Piero Andrea Temussi, 18 Dec 2020
Interactive discussion
-
SC1: 'Standards of MR', Gottfried Otting, 24 Oct 2020
- AC1: 'Otting observations', Piero Andrea Temussi, 29 Oct 2020
-
RC1: 'mr-2020-24', Anonymous Referee #1, 24 Oct 2020
-
AC2: 'reply to Anonymous observations', Piero Andrea Temussi, 29 Oct 2020
- AC3: 'revised answer to anonymous referee', Piero Andrea Temussi, 18 Dec 2020
-
AC2: 'reply to Anonymous observations', Piero Andrea Temussi, 29 Oct 2020
-
RC2: 'mr-2020-24', Dmitry Korzhnev, 27 Oct 2020
- AC4: 'answer to reviewer 2', Piero Andrea Temussi, 18 Dec 2020
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- 1
Rita Puglisi
Annalisa Pastore
Piero Andrea Temussi
This preprint has been withdrawn.
- Preprint
(817 KB) - Metadata XML
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Supplement
(275 KB) - BibTeX
- EndNote