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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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Formato: | Texto |
Lenguaje: | English |
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Public Library of Science
2007
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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. |
format | Text |
id | pubmed-2031916 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2007 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT vonderhaartobias optimizedproteinextractionforquantitativeproteomicsofyeasts |