Articolo in rivista, 2022, ENG, 10.1016/j.bpj.2022.03.010

Heat and cold denaturation of yeast frataxin: The effect of pressure

Puglisi Rita, Cioni Patrizia, Gabellieri Edi, Presciuttini Gianluca, Pastore Annalisa and Temussi Piero Andrea

Pastore, A (Corresponding Author), Kings Coll London, UK DRI Wohl Inst, London, England. Pastore, A (Corresponding Author), European Synchrotron Radiat Facil, Grenoble, France. Puglisi, Rita; Pastore, Annalisa; Temussi, Piero Andrea, Kings Coll London, UK DRI Wohl Inst, London, England. Cioni, Patrizia; Gabellieri, Edi; Presciuttini, Gianluca, CNR, Ist Biofis, Pisa, Italy. Pastore, Annalisa, European Synchrotron Radiat Facil, Grenoble, France.

Yfh1 is a yeast protein with the peculiar characteristic to undergo, in the absence of salt, cold denaturation at temperatures above the water freezing point. This feature makes the protein particularly interesting for studies aiming at understanding the rules that determine protein fold stability. Here, we present the phase diagram of Yfh1 unfolding as a function of pressure (0.1-500 MPa) and temperature 278-313 K (5-40 degrees C) both in the absence and in the presence of stabilizers using Trp fluorescence as a monitor. The protein showed a remarkable sensitivity to pressure: at 293 K, pressures around 10 MPa are sufficient to cause 50% of unfolding. Higher pressures were required for the unfolding of the protein in the presence of stabilizers. The phase diagram on the pressure-temperature plane together with a critical comparison between our results and those found in the literature allowed us to draw conclusions on the mechanism of the unfolding process under different environmental conditions.

Biophysical journal (Print) 121 (8), pp. 1502–1511

Keywords

FREE-ENERGY-LANDSCAPE, PROTEIN DENATURATION, STAPHYLOCOCCAL NUCLEASE, THERMAL-STABILITY, VOLUME CHANGE, IRON-BINDING, TEMPERATURE, HYDRATION, NMR, PHOSPHORESCENCE

CNR authors

Gabellieri Edi, Presciuttini Gianluca, Cioni Patrizia

CNR institutes

IBF – Istituto di biofisica

ID: 472732

Year: 2022

Type: Articolo in rivista

Creation: 2022-11-04 08:46:32.000

Last update: 2022-11-29 11:47:29.000

External links

OAI-PMH: Dublin Core

OAI-PMH: Mods

OAI-PMH: RDF

DOI: 10.1016/j.bpj.2022.03.010

External IDs

CNR OAI-PMH: oai:it.cnr:prodotti:472732

DOI: 10.1016/j.bpj.2022.03.010

ISI Web of Science (WOS): 000793038200014