Yeast frataxin is stabilized by low salt concentrations: cold denaturation disentangles ionic strength effects from specific interactions.

Frataxins are a family of metal binding proteins associated with the human Friedreich's ataxia disease. Here, we have addressed the effect of non-specifically binding salts on the stability of the yeast ortholog Yfh1. This protein is a sensitive model since its stability is strongly dependent o...

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Main Authors: Domenico Sanfelice, Rita Puglisi, Stephen R Martin, Lorenzo Di Bari, Annalisa Pastore, Piero Andrea Temussi
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2014-01-01
Series:PLoS ONE
Online Access:https://doi.org/10.1371/journal.pone.0095801
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author Domenico Sanfelice
Rita Puglisi
Stephen R Martin
Lorenzo Di Bari
Annalisa Pastore
Piero Andrea Temussi
author_facet Domenico Sanfelice
Rita Puglisi
Stephen R Martin
Lorenzo Di Bari
Annalisa Pastore
Piero Andrea Temussi
author_sort Domenico Sanfelice
collection DOAJ
description Frataxins are a family of metal binding proteins associated with the human Friedreich's ataxia disease. Here, we have addressed the effect of non-specifically binding salts on the stability of the yeast ortholog Yfh1. This protein is a sensitive model since its stability is strongly dependent on the environment, in particular on ionic strength. Yfh1 also offers the unique advantage that its cold denaturation can be observed above the freezing point of water, thus allowing the facile construction of the whole protein stability curve and hence the measurement of accurate thermodynamic parameters for unfolding. We systematically measured the effect of several cations and, as a control, of different anions. We show that, while strongly susceptible to ionic strength, as it would be in the cellular environment, Yfh1 stability is sensitive not only to divalent cations, which bind specifically, but also to monovalent cations. We pinpoint the structural bases of the stability and hypothesize that the destabilization induced by an unusual cluster of negatively charged residues favours the entrance of water molecules into the hydrophobic core, consistent with the generally accepted mechanism of cold denaturation.
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spelling doaj-art-b07dec8b4c014e6cb354bf3acef3fa662025-08-20T02:09:15ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0195e9580110.1371/journal.pone.0095801Yeast frataxin is stabilized by low salt concentrations: cold denaturation disentangles ionic strength effects from specific interactions.Domenico SanfeliceRita PuglisiStephen R MartinLorenzo Di BariAnnalisa PastorePiero Andrea TemussiFrataxins are a family of metal binding proteins associated with the human Friedreich's ataxia disease. Here, we have addressed the effect of non-specifically binding salts on the stability of the yeast ortholog Yfh1. This protein is a sensitive model since its stability is strongly dependent on the environment, in particular on ionic strength. Yfh1 also offers the unique advantage that its cold denaturation can be observed above the freezing point of water, thus allowing the facile construction of the whole protein stability curve and hence the measurement of accurate thermodynamic parameters for unfolding. We systematically measured the effect of several cations and, as a control, of different anions. We show that, while strongly susceptible to ionic strength, as it would be in the cellular environment, Yfh1 stability is sensitive not only to divalent cations, which bind specifically, but also to monovalent cations. We pinpoint the structural bases of the stability and hypothesize that the destabilization induced by an unusual cluster of negatively charged residues favours the entrance of water molecules into the hydrophobic core, consistent with the generally accepted mechanism of cold denaturation.https://doi.org/10.1371/journal.pone.0095801
spellingShingle Domenico Sanfelice
Rita Puglisi
Stephen R Martin
Lorenzo Di Bari
Annalisa Pastore
Piero Andrea Temussi
Yeast frataxin is stabilized by low salt concentrations: cold denaturation disentangles ionic strength effects from specific interactions.
PLoS ONE
title Yeast frataxin is stabilized by low salt concentrations: cold denaturation disentangles ionic strength effects from specific interactions.
title_full Yeast frataxin is stabilized by low salt concentrations: cold denaturation disentangles ionic strength effects from specific interactions.
title_fullStr Yeast frataxin is stabilized by low salt concentrations: cold denaturation disentangles ionic strength effects from specific interactions.
title_full_unstemmed Yeast frataxin is stabilized by low salt concentrations: cold denaturation disentangles ionic strength effects from specific interactions.
title_short Yeast frataxin is stabilized by low salt concentrations: cold denaturation disentangles ionic strength effects from specific interactions.
title_sort yeast frataxin is stabilized by low salt concentrations cold denaturation disentangles ionic strength effects from specific interactions
url https://doi.org/10.1371/journal.pone.0095801
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