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 proteins from yeast cells are likely to introduce artifacts into quantification procedures because of incomplete or selective extraction.


RESULTS
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.
von der Haar
Tobias
T
Protein Science Group, Department of Biosciences, University of Kent, Canterbury, United Kingdom. T.von-der-Haar@kent.ac.uk
D001671:Biochemistry / Q000379:methods*; D019295:Computational Biology; D005656:Fungal Proteins / Q000737:chemistry* / Q000302:isolation & purification; D015966:Gene Expression Regulation, Fungal; D008954:Models, Biological; D040081:Protein Array Analysis / Q000379:methods; D011506:Proteins / Q000737:chemistry; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D049490:Systems Biology; D015003:Yeasts
D005656:Fungal Proteins; D011506:Proteins; D020543:Proteome; D029701:Saccharomyces cerevisiae Proteins; D012967:Sodium Dodecyl Sulfate