Regulation of yeast central metabolism by enzyme phosphorylation
Abstract As a frequent post‐translational modification, protein phosphorylation regulates many cellular processes. Although several hundred phosphorylation sites have been mapped to metabolic enzymes in Saccharomyces cerevisiae, functionality was demonstrated for few of them. Here, we describe a nov...
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| Format: | Article |
| Language: | English |
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Springer Nature
2012-11-01
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| Series: | Molecular Systems Biology |
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| Online Access: | https://doi.org/10.1038/msb.2012.55 |
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| _version_ | 1850179510103703552 |
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| author | Ana Paula Oliveira Christina Ludwig Paola Picotti Maria Kogadeeva Ruedi Aebersold Uwe Sauer |
| author_facet | Ana Paula Oliveira Christina Ludwig Paola Picotti Maria Kogadeeva Ruedi Aebersold Uwe Sauer |
| author_sort | Ana Paula Oliveira |
| collection | DOAJ |
| description | Abstract As a frequent post‐translational modification, protein phosphorylation regulates many cellular processes. Although several hundred phosphorylation sites have been mapped to metabolic enzymes in Saccharomyces cerevisiae, functionality was demonstrated for few of them. Here, we describe a novel approach to identify in vivo functionality of enzyme phosphorylation by combining flux analysis with proteomics and phosphoproteomics. Focusing on the network of 204 enzymes that constitute the yeast central carbon and amino‐acid metabolism, we combined protein and phosphoprotein levels to identify 35 enzymes that change their degree of phosphorylation during growth under five conditions. Correlations between previously determined intracellular fluxes and phosphoprotein abundances provided first functional evidence for five novel phosphoregulated enzymes in this network, adding to nine known phosphoenzymes. For the pyruvate dehydrogenase complex E1 α subunit Pda1 and the newly identified phosphoregulated glycerol‐3‐phosphate dehydrogenase Gpd1 and phosphofructose‐1‐kinase complex β subunit Pfk2, we then validated functionality of specific phosphosites through absolute peptide quantification by targeted mass spectrometry, metabolomics and physiological flux analysis in mutants with genetically removed phosphosites. These results demonstrate the role of phosphorylation in controlling the metabolic flux realised by these three enzymes. |
| format | Article |
| id | doaj-art-eb7769db5d2c42d29b5d6afa8ba4dbe5 |
| institution | OA Journals |
| issn | 1744-4292 |
| language | English |
| publishDate | 2012-11-01 |
| publisher | Springer Nature |
| record_format | Article |
| series | Molecular Systems Biology |
| spelling | doaj-art-eb7769db5d2c42d29b5d6afa8ba4dbe52025-08-20T02:18:28ZengSpringer NatureMolecular Systems Biology1744-42922012-11-018111310.1038/msb.2012.55Regulation of yeast central metabolism by enzyme phosphorylationAna Paula Oliveira0Christina Ludwig1Paola Picotti2Maria Kogadeeva3Ruedi Aebersold4Uwe Sauer5Department of Biology, Institute of Molecular Systems Biology, ETH ZurichDepartment of Biology, Institute of Molecular Systems Biology, ETH ZurichDepartment of Biology, Institute of Molecular Systems Biology, ETH ZurichDepartment of Biology, Institute of Molecular Systems Biology, ETH ZurichDepartment of Biology, Institute of Molecular Systems Biology, ETH ZurichDepartment of Biology, Institute of Molecular Systems Biology, ETH ZurichAbstract As a frequent post‐translational modification, protein phosphorylation regulates many cellular processes. Although several hundred phosphorylation sites have been mapped to metabolic enzymes in Saccharomyces cerevisiae, functionality was demonstrated for few of them. Here, we describe a novel approach to identify in vivo functionality of enzyme phosphorylation by combining flux analysis with proteomics and phosphoproteomics. Focusing on the network of 204 enzymes that constitute the yeast central carbon and amino‐acid metabolism, we combined protein and phosphoprotein levels to identify 35 enzymes that change their degree of phosphorylation during growth under five conditions. Correlations between previously determined intracellular fluxes and phosphoprotein abundances provided first functional evidence for five novel phosphoregulated enzymes in this network, adding to nine known phosphoenzymes. For the pyruvate dehydrogenase complex E1 α subunit Pda1 and the newly identified phosphoregulated glycerol‐3‐phosphate dehydrogenase Gpd1 and phosphofructose‐1‐kinase complex β subunit Pfk2, we then validated functionality of specific phosphosites through absolute peptide quantification by targeted mass spectrometry, metabolomics and physiological flux analysis in mutants with genetically removed phosphosites. These results demonstrate the role of phosphorylation in controlling the metabolic flux realised by these three enzymes.https://doi.org/10.1038/msb.2012.55metabolic fluxmetabolismphosphoproteomepost‐translational regulationselected reaction monitoring |
| spellingShingle | Ana Paula Oliveira Christina Ludwig Paola Picotti Maria Kogadeeva Ruedi Aebersold Uwe Sauer Regulation of yeast central metabolism by enzyme phosphorylation Molecular Systems Biology metabolic flux metabolism phosphoproteome post‐translational regulation selected reaction monitoring |
| title | Regulation of yeast central metabolism by enzyme phosphorylation |
| title_full | Regulation of yeast central metabolism by enzyme phosphorylation |
| title_fullStr | Regulation of yeast central metabolism by enzyme phosphorylation |
| title_full_unstemmed | Regulation of yeast central metabolism by enzyme phosphorylation |
| title_short | Regulation of yeast central metabolism by enzyme phosphorylation |
| title_sort | regulation of yeast central metabolism by enzyme phosphorylation |
| topic | metabolic flux metabolism phosphoproteome post‐translational regulation selected reaction monitoring |
| url | https://doi.org/10.1038/msb.2012.55 |
| work_keys_str_mv | AT anapaulaoliveira regulationofyeastcentralmetabolismbyenzymephosphorylation AT christinaludwig regulationofyeastcentralmetabolismbyenzymephosphorylation AT paolapicotti regulationofyeastcentralmetabolismbyenzymephosphorylation AT mariakogadeeva regulationofyeastcentralmetabolismbyenzymephosphorylation AT ruediaebersold regulationofyeastcentralmetabolismbyenzymephosphorylation AT uwesauer regulationofyeastcentralmetabolismbyenzymephosphorylation |