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Dynamics of protein phosphorylation on Ser/Thr/Tyr in Bacillus subtilis
Author(s) -
Eymann Christine,
Becher Dörte,
Bernhardt Jörg,
Gronau Katrin,
Klutzny Anja,
Hecker Michael
Publication year - 2007
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200700232
Subject(s) - phosphorylation , bacillus subtilis , threonine , protein phosphorylation , dephosphorylation , biochemistry , serine , biology , phosphoserine , chemistry , phosphatase , protein kinase a , bacteria , genetics
The Ser/Thr/Tyr phosphoproteome of Bacillus subtilis was analyzed by a 2‐D gel‐based approach combining Pro‐Q Diamond staining and [ 33 P]‐labeling. In exponentially growing B. subtilis cells 27 proteins could be identified after staining with Pro‐Q Diamond and/or [ 33 P]‐labeling and one additional protein was labeled solely by [ 33 P] resulting in a total of 28 potentially phosphorylated proteins. These proteins are mainly involved in enzymatic reactions of basic carbon metabolism and the regulation of the alternative sigma factor σ B . We also found significant changes of the phosphoproteome including increased phosphorylation and dephosphorylation rates of some proteins as well as the detection of four newly phosphorylated proteins in response to stress or starvation. For nine proteins, phosphorylation sites at serine or threonine residues were determined by MS. These include the known phosphorylation sites of Crh, PtsH, and RsbV. Additionally, we were able to identify novel phosphorylation sites of AroA, Pyk, and YbbT. Interestingly, the phosphorylation of RsbRA, B, C, and D, four proteins of a multicomponent protein complex involved in environmental stress signaling, was found during exponential growth. For RsbRA, B, and D, phosphorylation of one of the conserved threonine residues in their C‐termini were verified by MS (T171, T186, T181, respectively).