La maladie de Parkinson en France (serveur d'exploration)

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Qualitative and Quantitative Multiplexed Proteomic Analysis of Complex Yeast Protein Fractions That Modulate the Assembly of the Yeast Prion Sup35p

Identifieur interne : 001618 ( Main/Merge ); précédent : 001617; suivant : 001619

Qualitative and Quantitative Multiplexed Proteomic Analysis of Complex Yeast Protein Fractions That Modulate the Assembly of the Yeast Prion Sup35p

Auteurs : Virginie Redeker [France] ; Chris Hughes [Royaume-Uni] ; Jimmy Savistchenko [France] ; Johannes P. C. Vissers [Royaume-Uni] ; Ronald Melki [France]

Source :

RBID : PMC:3172207

Abstract

Background

The aggregation of the baker's yeast prion Sup35p is at the origin of the transmissible [PSI+] trait. We and others have shown that molecular chaperones modulate Sup35p aggregation. However, other protein classes might be involved in [PSI+] formation.

Results

We designed a functional proteomic study that combines two techniques to identify modulators of Sup35p aggregation and describe the changes associated to [PSI+] formation. The first allows measuring the effect of fractionated Saccharomyces cerevisiae cytosolic extracts from [PSI+] and [psi] yeast cells on Sup35p assembly. The second is a multiplex qualitative and quantitative comparison of protein composition of active and inactive fractions using a gel-free and label-free LC-MS approach. We identify changes in proteins involved in translation, folding, degradation, oxido-reduction and metabolic processes.

Conclusion

Our functional proteomic study provides the first inventory list of over 300 proteins that directly or indirectly affect Sup35p aggregation and [PSI+] formation. Our results highlight the complexity of the cellular changes accompanying [PSI+] formation and pave the way for in vitro studies aimed to document the effect of individual and/or combinations of proteins identified here, susceptible of affecting Sup35p assembly.


Url:
DOI: 10.1371/journal.pone.0023659
PubMed: 21931608
PubMed Central: 3172207

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PMC:3172207

Le document en format XML

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<title xml:lang="en" level="a" type="main">Qualitative and Quantitative Multiplexed Proteomic Analysis of Complex Yeast Protein Fractions That Modulate the Assembly of the Yeast Prion Sup35p</title>
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<title>Background</title>
<p>The aggregation of the baker's yeast prion Sup35p is at the origin of the transmissible [
<italic>PSI
<sup>+</sup>
</italic>
] trait. We and others have shown that molecular chaperones modulate Sup35p aggregation. However, other protein classes might be involved in [
<italic>PSI
<sup>+</sup>
</italic>
] formation.</p>
</sec>
<sec>
<title>Results</title>
<p>We designed a functional proteomic study that combines two techniques to identify modulators of Sup35p aggregation and describe the changes associated to [
<italic>PSI
<sup>+</sup>
</italic>
] formation. The first allows measuring the effect of fractionated
<italic>Saccharomyces cerevisiae</italic>
cytosolic extracts from [
<italic>PSI
<sup>+</sup>
</italic>
] and [
<italic>psi
<sup></sup>
</italic>
] yeast cells on Sup35p assembly. The second is a multiplex qualitative and quantitative comparison of protein composition of active and inactive fractions using a gel-free and label-free LC-MS approach. We identify changes in proteins involved in translation, folding, degradation, oxido-reduction and metabolic processes.</p>
</sec>
<sec>
<title>Conclusion</title>
<p>Our functional proteomic study provides the first inventory list of over 300 proteins that directly or indirectly affect Sup35p aggregation and [
<italic>PSI
<sup>+</sup>
</italic>
] formation. Our results highlight the complexity of the cellular changes accompanying [
<italic>PSI
<sup>+</sup>
</italic>
] formation and pave the way for
<italic>in vitro</italic>
studies aimed to document the effect of individual and/or combinations of proteins identified here, susceptible of affecting Sup35p assembly.</p>
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