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The anatomy of unfolding of Yfh1 is revealed by site-specific fold stability analysis measured by 2D NMR spectroscopy.
Puglisi, Rita; Karunanithy, Gogulan; Hansen, D Flemming; Pastore, Annalisa; Temussi, Piero Andrea.
Afiliación
  • Puglisi R; UK-DRI at King's College London, The Wohl Institute, London, UK.
  • Karunanithy G; Department of Structural Biology, Division of Biosciences, University College London, London, UK.
  • Hansen DF; Department of Structural Biology, Division of Biosciences, University College London, London, UK.
  • Pastore A; UK-DRI at King's College London, The Wohl Institute, London, UK.
  • Temussi PA; European Synchrotron Radiation Facility, Grenoble, France.
Commun Chem ; 4(1)2021 Dec.
Article en En | MEDLINE | ID: mdl-35243007
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 an 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.

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Commun Chem Año: 2021 Tipo del documento: Article Pais de publicación: Reino Unido

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Commun Chem Año: 2021 Tipo del documento: Article Pais de publicación: Reino Unido