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Optimized Protein Extraction for Quantitative Proteomics of Yeasts

BACKGROUND: The absolute quantification of intracellular protein levels is technically demanding, but has recently become more prominent because novel approaches like systems biology and metabolic control analysis require knowledge of these parameters. Current protocols for the extraction of protein...

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Detalles Bibliográficos
Autor principal: von der Haar, Tobias
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2007
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2031916/
https://www.ncbi.nlm.nih.gov/pubmed/17957260
http://dx.doi.org/10.1371/journal.pone.0001078
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author von der Haar, Tobias
author_facet von der Haar, Tobias
author_sort von der Haar, Tobias
collection PubMed
description BACKGROUND: The absolute quantification of intracellular protein levels is technically demanding, but has recently become more prominent because novel approaches like systems biology and metabolic control analysis require knowledge of these parameters. Current protocols for the extraction of proteins from yeast cells are likely to introduce artifacts into quantification procedures because of incomplete or selective extraction. PRINCIPAL FINDINGS: We have developed a novel procedure for protein extraction from S. cerevisiae based on chemical lysis and simultaneous solubilization in SDS and urea, which can extract the great majority of proteins to apparent completeness. The procedure can be used for different Saccharomycetes yeast species and varying growth conditions, is suitable for high-throughput extraction in a 96-well format, and the resulting extracts can easily be post-processed for use in non-SDS compatible procedures like 2D gel electrophoresis. CONCLUSIONS: An improved method for quantitative protein extraction has been developed that removes some of the sources of artefacts in quantitative proteomics experiments, while at the same time allowing novel types of applications.
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spelling pubmed-20319162007-10-24 Optimized Protein Extraction for Quantitative Proteomics of Yeasts von der Haar, Tobias PLoS One Research Article BACKGROUND: The absolute quantification of intracellular protein levels is technically demanding, but has recently become more prominent because novel approaches like systems biology and metabolic control analysis require knowledge of these parameters. Current protocols for the extraction of proteins from yeast cells are likely to introduce artifacts into quantification procedures because of incomplete or selective extraction. PRINCIPAL FINDINGS: We have developed a novel procedure for protein extraction from S. cerevisiae based on chemical lysis and simultaneous solubilization in SDS and urea, which can extract the great majority of proteins to apparent completeness. The procedure can be used for different Saccharomycetes yeast species and varying growth conditions, is suitable for high-throughput extraction in a 96-well format, and the resulting extracts can easily be post-processed for use in non-SDS compatible procedures like 2D gel electrophoresis. CONCLUSIONS: An improved method for quantitative protein extraction has been developed that removes some of the sources of artefacts in quantitative proteomics experiments, while at the same time allowing novel types of applications. Public Library of Science 2007-10-24 /pmc/articles/PMC2031916/ /pubmed/17957260 http://dx.doi.org/10.1371/journal.pone.0001078 Text en Tobias von der Haar. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
von der Haar, Tobias
Optimized Protein Extraction for Quantitative Proteomics of Yeasts
title Optimized Protein Extraction for Quantitative Proteomics of Yeasts
title_full Optimized Protein Extraction for Quantitative Proteomics of Yeasts
title_fullStr Optimized Protein Extraction for Quantitative Proteomics of Yeasts
title_full_unstemmed Optimized Protein Extraction for Quantitative Proteomics of Yeasts
title_short Optimized Protein Extraction for Quantitative Proteomics of Yeasts
title_sort optimized protein extraction for quantitative proteomics of yeasts
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2031916/
https://www.ncbi.nlm.nih.gov/pubmed/17957260
http://dx.doi.org/10.1371/journal.pone.0001078
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