Phosphoproteome analysis of the pathogenic bacterium Helicobacter pylori reveals over-representation of tyrosine phosphorylation and multiply phosphorylated proteins

• Published in: Proteomics (Weinheim) Vol. 11; no. 8; pp. 1449 - 1461
• Main Authors: Ge, Ruiguang, Sun, Xuesong, Xiao, Chuanle, Yin, Xingfeng, Shan, Weiran, Chen, Zhuo, He, Qing-Yu
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 01.04.2011; WILEY‐VCH Verlag; Wiley-VCH; Wiley Subscription Services, Inc
• Subjects: Analytical, structural and metabolic biochemistry; Bacteria; Bacterial diseases; Bacterial diseases of the digestive system and abdomen; Bacterial Proteins - analysis; Bacterial Proteins - chemistry; Biological and medical sciences; Chromatography, Liquid; Fundamental and applied biological sciences. Psychology; Helicobacter pylori; Helicobacter pylori - chemistry; Helicobacter pylori - metabolism; Human bacterial diseases; Infectious diseases; Kinases; Mass Spectrometry; Medical sciences; Membrane; Microbiology; Miscellaneous; Phosphopeptides - analysis; Phosphopeptides - metabolism; Phosphoproteins - analysis; Phosphoproteins - metabolism; Phosphoproteome; Phosphorylation; Phosphotyrosine - analysis; Phosphotyrosine - metabolism; Protein Kinases - metabolism; Proteins; Proteome - analysis; Proteome - metabolism; Proteomics; Tyrosine phosphorylation; Ulcers; Tyrosine; Phosphorylation; Spirillales; Microbiology; Spirillaceae; Protein; Helicobacter pylori; Tyrosine phosphorylation; Pathogenic; Proteomics; Bacteria; Membrane; Phosphoproteome
• ISSN: 1615-9853; 1615-9861; 1615-9861
• DOI: 10.1002/pmic.201000649

Increasing evidence shows that protein phosphorylation on serine (Ser), threonine (Thr) and tyrosine (Tyr) residues is a major regulatory post‐translational modification in the bacteria. To reveal the phosphorylation state in the Gram‐negative pathogenic bacterium Helicobacter pylori, we carried out a global and site‐specific phosphoproteomic analysis based on TiO2‐phosphopeptide enrichment and high‐accuracy LC‐MS/MS determination. Eighty‐two phosphopeptides from 67 proteins were identified with 126 phosphorylation sites, among which 79 class I sites were determined to have a distribution of 42.8:38.7:18.5% for the Ser/Thr/Tyr phosphorylation, respectively. The H. pylori phosphoproteome is characterized by comparably big size, high ratio of Tyr phosphorylation, high abundance of multiple phosphorylation sites in individual phosphopeptides and over‐representation of membrane proteins. An interaction network covering 28 phosphoproteins was constructed with a total of 163 proteins centering on the major H. pylori virulence factor VacA, indicating that protein phosphorylation in H. pylori may be delicately controlled to regulate many aspects of the metabolic pathways and bacterial virulence.