Magnetic Resonance
Magnetic Resonance
Magnetic Resonance
Preprints
https://doi.org/10.5194/mr-2020-24
https://doi.org/10.5194/mr-2020-24

  07 Oct 2020

07 Oct 2020

Review status: a revised version of this preprint is currently under review for the journal MR.

Anatomy of unfolding: The site-specific fold stability of Yfh1 measured by 2D NMR

Rita Puglisi, Annalisa Pastore, and Piero Andrea Temussi Rita Puglisi et al.
  • UK-DRI at King's College London, The Wohl Institute, 5 Cutcombe Rd, SE59RT London, UK

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.

Rita Puglisi et al.

 
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Rita Puglisi et al.

Rita Puglisi et al.

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Short summary
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. We asked whether a detailed analysis of the individual stability curves could provide additional site-specific information. By monitoring both cold and heat denaturation of individual residues we convincingly demonstrate that the two unfolding processes are intrinsically different.