Ser/Thr/Tyr phosphoproteome analysis of pathogenic and non-pathogenic Pseudomonas species.
Protein phosphorylation on serine, threonine, and tyrosine is well established as a crucial regulatory posttranslational modification in eukaryotes. With the recent whole-genome sequencing projects reporting the presence of serine/threonine kinases and two-component proteins both in prokaryotes and eukaryotes, the importance of protein phosphorylation in archaea and bacteria is gaining acceptance. While conventional biochemical methods failed to obtain a snapshot of the bacterial phosphoproteomes, advances in MS methods have paved the way for in-depth mapping of phosphorylation sites. Here, we present phosphoproteomes of two ecologically diverse non-enteric Gram-negative bacteria captured by a nanoLC-MS-based approach combined with a novel phosphoenrichment method. While the phosphoproteome data from the two species are not very similar, the results reflect high similarity to the previously published dataset in terms of the pathways the phosphoproteins belong to. This study additionally provides evidence to prior observations that protein phosphorylation is common in bacteria. Notably, phosphoproteins identified in Pseudomonas aeruginosa belong to motility, transport, and pathogenicity pathways that are critical for survival and virulence. We report, for the first time, that motility regulator A, probably acting via the novel secondary messenger cyclic diguanylate monophosphate, significantly affects protein phosphorylation in Pseudomonas putida.
Ravichandran
Ayshwarya
A
Department of Biological Sciences, National University of Singapore, 10 Science Drive 4, Singapore.
Sugiyama
Naoyuki
N
Tomita
Masaru
M
Swarup
Sanjay
S
Ishihama
Yasushi
Y
D000596:Amino Acids / Q000378:metabolism*; D001426:Bacterial Proteins / Q000032:analysis* / Q000378:metabolism; D002853:Chromatography, Liquid; D006152:Cyclic GMP / Q000031:analogs & derivatives / Q000378:metabolism; D013058:Mass Spectrometry; D019606:Phosphoamino Acids / Q000378:metabolism; D010748:Phosphopeptides / Q000032:analysis* / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D020543:Proteome / Q000032:analysis* / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000737:chemistry* / Q000378:metabolism; D016958:Pseudomonas putida / Q000737:chemistry* / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D015398:Signal Transduction; D013912:Threonine / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism
D000596:Amino Acids; D001426:Bacterial Proteins; D019606:Phosphoamino Acids; D010748:Phosphopeptides; D020543:Proteome; D013912:Threonine; D014443:Tyrosine; D012694:Serine; C062025:bis(3',5')-cyclic diguanylic acid; D011494:Protein Kinases; D006152:Cyclic GMP