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Quantitative Phosph...
Quantitative Phosphoproteome Analysis of Bacillus subtilis Reveals Novel Substrates of the Kinase PrkC and Phosphatase PrpC
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- Ravikumar, V. (author)
- Eberhard Karls Universität Tübingen,Eberhard Karls University of Tübingen
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Shi, L. (author)
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- Krug, K. (author)
- Eberhard Karls Universität Tübingen,Eberhard Karls University of Tübingen
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Derouiche, A. (author)
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Jers, C. (author)
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Cousin, C. (author)
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Kobir, A. (author)
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- Mijakovic, Ivan, 1975 (author)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Macek, B. (author)
- Eberhard Karls Universität Tübingen,Eberhard Karls University of Tübingen
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(creator_code:org_t)
- 2014
- 2014
- English.
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In: Molecular and Cellular Proteomics. - 1535-9476 .- 1535-9484. ; 13:8, s. 1965-1978
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Abstract
Subject headings
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- Reversible protein phosphorylation on serine, threonine, and tyrosine (Ser/Thr/Tyr) residues plays a critical role in regulation of vital processes in the cell. Despite of considerable progress in our understanding of the role of this modification in bacterial physiology, the dynamics of protein phosphorylation during bacterial growth has rarely been systematically addressed. In addition, little is known about in vivo substrates of bacterial Ser/Thr/Tyr kinases and phosphatases. An excellent candidate to study these questions is the Gram-positive bacterium Bacillus subtilis, one of the most intensively investigated bacterial model organism with both research and industrial applications. Here we employed gel-free phosphoproteomics combined with SILAC labeling and high resolution mass spectrometry to study the proteome and phosphoproteome dynamics during the batch growth of B. subtilis. We measured the dynamics of 1666 proteins and 64 phosphorylation sites in five distinct phases of growth. Enzymes of the central carbon metabolism and components of the translation machinery appear to be highly phosphorylated in the stationary phase, coinciding with stronger expression of Ser/Thr kinases. We further used the SILAC workflow to identify novel putative substrates of the Ser/Thr kinase PrkC and the phosphatase PrpC during stationary phase. The overall number of putative substrates was low, pointing to a high kinase and phosphatase specificity. One of the phosphorylation sites affected by both, PrkC and PrpC, was the Ser281 on the oxidoreductase YkwC. We showed that PrkC phosphorylates and PrpC dephosphorylates YkwC in vitro and that phosphorylation at Ser281 abolishes the oxidoreductase activity of YkwC in vitro and in vivo. Our results present the most detailed phosphoproteomic analysis of B. subtilis growth to date and provide the first global in vivo screen of PrkC and PrpC substrates.
Subject headings
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Keyword
- ESCHERICHIA-COLI
- AMINO-ACIDS
- GRAM-POSITIVE BACTERIA
- TYROSINE PHOSPHORYLATION
- PROTEIN-KINASE
- PROTEOMICS
- SER/THR/TYR PHOSPHOPROTEOME
- MEMBRANE
- SERINE/THREONINE
- LACTOCOCCUS-LACTIS
Publication and Content Type
- art (subject category)
- ref (subject category)
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- By the author/editor
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Ravikumar, V.
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Shi, L.
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Krug, K.
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Derouiche, A.
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Jers, C.
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Cousin, C.
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show more...
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Kobir, A.
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Mijakovic, Ivan, ...
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Macek, B.
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show less...
- About the subject
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
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and Biochemistry and ...
- Articles in the publication
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Molecular and Ce ...
- By the university
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Chalmers University of Technology