Endogenous occurrence of protein S-guanylation in Escherichia coli: Target identification and genetic regulation

Hiroyasu Tsutsuki, Minkyung Jung, Tianli Zhang, Katsuhiko Ono, Tomoaki Ida, Kohei Kunieda, Hideshi Ihara, Takaaki Akaike, Tomohiro Sawa

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)


8-Nitroguanosine 3′,5′-cyclic monophosphate (8-nitro-cGMP) is a nitrated cGMP derivative formed in response to nitric oxide (NO) and reactive oxygen species (ROS). It can cause a post-translational modification (PTM) of protein thiols through cGMP adduction (protein S-guanylation). Accumulating evidence has suggested that, in mammals, S-guanylation of redox-sensor proteins may implicate in regulation of adaptive responses against ROS-associated oxidative stress. Occurrence as well as protein targets of S-guanylation in bacteria remained unknown, however. Here we demonstrated, for the first time, the endogenous occurrence of protein S-guanylation in Escherichia coli (E. coli). Western blotting using anti-S-guanylation antibody clearly showed that multiple proteins were S-guanylated in E. coli. Interestingly, some of those proteins were more intensely S-guanylated when bacteria were cultured under static culture condition than shaking culture condition. It has been known that E. coli is deficient of guanylate cyclase, an enzyme indispensable for 8-nitro-cGMP formation in mammals. We found that adenylate cyclase from E. coli potentially catalyzed 8-nitro-cGMP formation from its precursor 8-nitroguanosine 5′-triphosphate. More importantly, E. coli lacking adenylate cyclase showed significantly reduced formation of S-guanylated proteins. Our S-guanylation proteomics successfully identified S-guanylation protein targets in E. coli, including chaperons, ribosomal proteins, and enzymes which associate with protein synthesis, redox regulation and metabolism. Understanding of functional impacts for protein S-guanylation in bacterial signal transduction is necessary basis for development of potential chemotherapy and new diagnostic strategy for control of pathogenic bacterial infections.

Original languageEnglish
Pages (from-to)7-11
Number of pages5
JournalBiochemical and biophysical research communications
Issue number1
Publication statusPublished - 2016 Sept 9


  • 8-Nitro-cGMP
  • Adenylate cyclase
  • Escherichia coli
  • Nitric oxide
  • Protein S-guanylation
  • Redox signaling

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology


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