von der Haar, Tobias (2007) Optimized protein extraction for quantitative proteomics of yeasts. PLoS ONE, 2 (10). e1078. ISSN 1932-6203. (doi:https://doi.org/10.1371/journal.pone.0001078) (Full text available)
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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 Saccharomyces 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.
|Uncontrolled keywords:||Yeast, Saccharomyces cerevisiae, protein chemistry, proteomics, eRF3, Sup35|
Faculties > Sciences > School of Biosciences
Faculties > Sciences > School of Biosciences > Protein Science Group
|Depositing User:||Tobias von der Haar|
|Date Deposited:||24 Jul 2008 09:40 UTC|
|Last Modified:||07 May 2014 08:24 UTC|
|Resource URI:||https://kar.kent.ac.uk/id/eprint/3949 (The current URI for this page, for reference purposes)|