Direct Iterative Protein Profiling (DIPP) - an Innovative Method for Large-scale Protein Detection Applied to Budding Yeast Mitosis
The budding yeast Saccharomyces cerevisiae is a major model organism for important biological processes such as mitotic growth and meiotic development, it can be a human pathogen, and it is widely used in the food-, and biotechnology industries. Consequently, the genomes of numerous strains have bee...
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| Veröffentlicht in: | Molecular & cellular proteomics 2012-02, Vol.11 (2), p.M111.012682-M111.012682, Article M111.012682 |
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| container_issue | 2 |
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| container_title | Molecular & cellular proteomics |
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| creator | Lavigne, Régis Becker, Emmanuelle Liu, Yuchen Evrard, Bertrand Lardenois, Aurélie Primig, Michael Pineau, Charles |
| description | The budding yeast Saccharomyces cerevisiae is a major model organism for important biological processes such as mitotic growth and meiotic development, it can be a human pathogen, and it is widely used in the food-, and biotechnology industries. Consequently, the genomes of numerous strains have been sequenced and a very large amount of RNA profiling data is available. Moreover, it has recently become possible to quantitatively analyze the entire yeast proteome; however, efficient and cost-effective high-throughput protein profiling remains a challenge. We report here a new approach to direct and label-free large-scale yeast protein identification using a tandem buffer system for protein extraction, two-step protein prefractionation and enzymatic digestion, and detection of peptides by iterative mass spectrometry. Our profiling study of diploid cells undergoing rapid mitotic growth identified 86% of the known proteins and its output was found to be widely concordant with genome-wide mRNA concentrations and DNA variations between yeast strains. This paves the way for comprehensive and straightforward yeast proteome profiling across a wide variety of experimental conditions. |
| doi_str_mv | 10.1074/mcp.M111.012682 |
| format | Article |
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Consequently, the genomes of numerous strains have been sequenced and a very large amount of RNA profiling data is available. Moreover, it has recently become possible to quantitatively analyze the entire yeast proteome; however, efficient and cost-effective high-throughput protein profiling remains a challenge. We report here a new approach to direct and label-free large-scale yeast protein identification using a tandem buffer system for protein extraction, two-step protein prefractionation and enzymatic digestion, and detection of peptides by iterative mass spectrometry. Our profiling study of diploid cells undergoing rapid mitotic growth identified 86% of the known proteins and its output was found to be widely concordant with genome-wide mRNA concentrations and DNA variations between yeast strains. 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| ispartof | Molecular & cellular proteomics, 2012-02, Vol.11 (2), p.M111.012682-M111.012682, Article M111.012682 |
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| subjects | Biochemistry, Molecular Biology Chromatography, Liquid Data processing Diploids DNA DNA, Fungal - analysis DNA, Fungal - genetics Food industry Gene Expression Profiling Genomes Humans Life Sciences Mass spectroscopy Meiosis Mitosis Mitosis - physiology mRNA Pathogens Proteome - analysis Proteomics Saccharomyces cerevisiae Saccharomyces cerevisiae - growth & development Saccharomyces cerevisiae - metabolism Saccharomyces cerevisiae Proteins - genetics Saccharomyces cerevisiae Proteins - metabolism Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Technical Innovation and Resources |
| title | Direct Iterative Protein Profiling (DIPP) - an Innovative Method for Large-scale Protein Detection Applied to Budding Yeast Mitosis |
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