Unseen Proteome: Mining Below the Tip of the Iceberg To Find Low Abundance and Membrane Proteins
Abundant and hydrophilic nonmembrane proteins with isoelectric points below pH 8 are the predominant proteins identified in most proteomics projects. In yeast, however, low-abundance proteins make up 80% of the predicted proteome, approximately 50% have pI's above pH 8 and 30% of the yeast ORFs...
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| Veröffentlicht in: | Journal of proteome research 2003-05, Vol.2 (3), p.303-311 |
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| container_title | Journal of proteome research |
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| creator | Pedersen, Susanne K Harry, Jenny L Sebastian, Lucille Baker, Jasmine Traini, Mathew D McCarthy, John T Manoharan, Abi Wilkins, Marc R Gooley, Andrew A Righetti, Pier Giorgio Packer, Nicolle H Williams, Keith L Herbert, Ben R |
| description | Abundant and hydrophilic nonmembrane proteins with isoelectric points below pH 8 are the predominant proteins identified in most proteomics projects. In yeast, however, low-abundance proteins make up 80% of the predicted proteome, approximately 50% have pI's above pH 8 and 30% of the yeast ORFs are predicted to encode membrane proteins with at least 1 trans-membrane span. By applying highly solubilizing reagents and isoelectric fractionation to a membrane fraction of yeast we have a purified and identified 780 protein isoforms, representing 323 gene products, including 28% low abundance proteins and 49% membrane or membrane associated proteins. More importantly, considering the frequency and importance of co- and post-translational modifications, the separation of protein isoforms is essential and two-dimensional electrophoresis remains the only technique which offers sufficient resolution to address this at a proteomic level. Keywords: two-dimensional electrophoresis • membrane proteins • low-abundance proteins • multi-compartment electrolyzer • fractionation |
| doi_str_mv | 10.1021/pr025588i |
| format | Article |
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In yeast, however, low-abundance proteins make up 80% of the predicted proteome, approximately 50% have pI's above pH 8 and 30% of the yeast ORFs are predicted to encode membrane proteins with at least 1 trans-membrane span. By applying highly solubilizing reagents and isoelectric fractionation to a membrane fraction of yeast we have a purified and identified 780 protein isoforms, representing 323 gene products, including 28% low abundance proteins and 49% membrane or membrane associated proteins. More importantly, considering the frequency and importance of co- and post-translational modifications, the separation of protein isoforms is essential and two-dimensional electrophoresis remains the only technique which offers sufficient resolution to address this at a proteomic level. 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| subjects | Amino Acid Sequence Cytochrome-B Reductase - genetics Electrophoresis, Gel, Two-Dimensional - methods Hydrophobic and Hydrophilic Interactions Membrane Proteins - genetics Membrane Proteins - physiology Molecular Sequence Data Proteomics - methods Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae - physiology |
| title | Unseen Proteome: Mining Below the Tip of the Iceberg To Find Low Abundance and Membrane Proteins |
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