title: The phosphoproteome of the minimal bacterium Mycoplasma pneumoniae: analysis of the complete known Ser/Thr kinome suggests the existence of novel kinases.
abstract: Mycoplasma pneumoniae belongs to the Mollicutes, the group of organisms with the smallest genomes that are capable of host-independent life. These bacteria show little regulation in gene expression, suggesting an important role for the control of protein activities. We have studied protein phosphorylation in M. pneumoniae to identify phosphorylated proteins. Two-dimensional gel electrophoresis and mass spectrometry allowed the detection of 63 phosphorylated proteins, many of them enzymes of central carbon metabolism and proteins related to host cell adhesion. We identified 16 phosphorylation sites, among them 8 serine and 8 threonine residues, respectively. A phosphoproteome analysis with mutants affected in the two annotated protein kinase genes or in the single known protein phosphatase gene suggested that only one protein (HPr) is phosphorylated by the HPr kinase, HPrK, whereas four adhesion-related or surface proteins were targets of the protein kinase C, PrkC. A comparison with the phosphoproteomes of other bacteria revealed that protein phosphorylation is evolutionarily only poorly conserved. Only one single protein with an identified phosphorylation site, a phosphosugar mutase (ManB in M. pneumoniae), is phosphorylated on a conserved serine residue in all studied organisms from archaea and bacteria to man. We demonstrate that this protein undergoes autophosphorylation. This explains the strong conservation of this phosphorylation event. For most other proteins, even if they are phosphorylated in different species, the actual phosphorylation sites are different. This suggests that protein phosphorylation is a form of adaptation of the bacteria to the specific needs of their particular ecological niche.
authors.affiliation: Department of General Microbiology, Georg-August-Universität Göttingen, Grisebachstrasse 8, D-37077 Göttingen, Germany.
authors.forename: Sebastian R
authors.initials: SR
authors.lastname: Schmidl
authors.forename: Katrin
authors.initials: K
authors.lastname: Gronau
authors.forename: Nico
authors.initials: N
authors.lastname: Pietack
authors.forename: Michael
authors.initials: M
authors.lastname: Hecker
authors.forename: Dörte
authors.initials: D
authors.lastname: Becher
authors.forename: Jörg
authors.initials: J
authors.lastname: Stülke
mesh_terms: D000595:Amino Acid Sequence; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D017124:Conserved Sequence; D004789:Enzyme Activation; D006019:Glycolysis; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009177:Mycoplasma pneumoniae / Q000166:cytology / Q000201:enzymology* / Q000254:growth & development; D010750:Phosphoproteins / Q000737:chemistry / Q000145:classification / Q000378:metabolism*; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism*; D012694:Serine / Q000378:metabolism*; D013912:Threonine / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D010750:Phosphoproteins; D020543:Proteome; D013912:Threonine; D012694:Serine; D017346:Protein Serine-Threonine Kinases