The phosphoproteome and its physiological dynamics in Staphylococcus aureus.
Phosphorylation events on proteins during growth and stress/starvation can represent crucial regulation processes inside the bacterial cell. Therefore, serine, threonine and tyrosine phosphorylation patterns were analyzed by two powerful complementary proteomic methods for the human pathogen Staphylococcus aureus. Using 2D-gel analysis with a phosphosensitive stain (Pro-Q Diamond) and gel-free titanium dioxide based phosphopeptide enrichment, 103 putative phosphorylated proteins with successfully mapped 68 different phosphorylation sites were found in the soluble proteome of S. aureus. Additionally, in a proof of concept study, 8 proteins phosphorylated on arginine residues have been identified. Most important for functional analyses of S. aureus, proteins related to pathogenicity and virulence were found to be phosphorylated: the virulence regulator SarA, the potential antimicrobial target FbaA and the elastin-binding protein EbpS. Besides newly identified phosphorylation sites we compared our dataset with existing data from literature and subsequent experiments revealed additional phosphorylation events on highly conserved localizations in FbaA. Differential analysis of phosphorylation signals on the 2D-gels showed significant changes in phosphorylation under different physiological conditions for 10 proteins. Among these, we were able to detect newly appearing signals for phosphorylated isoforms of FdaB and HchA under nitrosative stress conditions.
Institute for Microbiology, Ernst Moritz Arndt University Greifswald, Germany.
Katrin
K
Bäsell
Institute for Microbiology, Ernst Moritz Arndt University Greifswald, Germany.
Andreas
A
Otto
Institute for Microbiology, Ernst Moritz Arndt University Greifswald, Germany.
Sabryna
S
Junker
Institute for Microbiology, Ernst Moritz Arndt University Greifswald, Germany.
Daniela
D
Zühlke
Institute for Microbiology, Ernst Moritz Arndt University Greifswald, Germany.
Gerd-Martin
GM
Rappen
Institute for Microbiology, Ernst Moritz Arndt University Greifswald, Germany.
Sabrina
S
Schmidt
Institute for Microbiology, Ernst Moritz Arndt University Greifswald, Germany.
Christian
C
Hentschker
Proteome Center, Interfaculty Institute for Cell Biology, University of Tübingen, Germany.
Boris
B
Macek
Institute for Molecular Infection Biology, Julius Maximillians University, Würzburg, Germany.
Knut
K
Ohlsen
Institute for Microbiology, Ernst Moritz Arndt University Greifswald, Germany.
Michael
M
Hecker
Institute for Microbiology, Ernst Moritz Arndt University Greifswald, Germany. Electronic address: dbecher@uni-greifswald.de.
Dörte
D
Becher
D000222:Adaptation, Physiological; D001426:Bacterial Proteins / Q000032:analysis*; D015180:Electrophoresis, Gel, Two-Dimensional; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D013058:Mass Spectrometry; D010750:Phosphoproteins / Q000032:analysis*; D020543:Proteome / Q000032:analysis*; D013194:Staining and Labeling; D013211:Staphylococcus aureus / Q000737:chemistry*
D001426:Bacterial Proteins; D010750:Phosphoproteins; D020543:Proteome
2D-gel; Arginine/serine/threonine/tyrosine phosphorylation; Phosphopeptide enrichment; Phosphoproteome; Pro-Q diamond; Staphylococcus aureus