title: Persistent Activation of cGMP-Dependent Protein Kinase by a Nitrated Cyclic Nucleotide via Site Specific Protein S-Guanylation.
abstract: 8-Nitroguanosine 3',5'-cyclic monophosphate (8-nitro-cGMP) is a nitrated derivative of guanosine 3',5'-cyclic monophosphate (cGMP) formed endogenously under conditions associated with production of both reactive oxygen species and nitric oxide. It acts as an electrophilic second messenger in the regulation of cellular signaling by inducing a post-translational modification of redox-sensitive protein thiols via covalent adduction of cGMP moieties to protein thiols (protein S-guanylation). Here, we demonstrate that 8-nitro-cGMP potentially S-guanylates thiol groups of cGMP-dependent protein kinase (PKG), the enzyme that serves as one of the major receptor proteins for intracellular cGMP and controls a variety of cellular responses. S-Guanylation of PKG was found to occur in a site specific manner; Cys42 and Cys195 were the susceptible residues among 11 Cys residues. Importantly, S-guanylation at Cys195, which is located in the high-affinity cGMP binding domain of PKG, causes persistent enzyme activation as determined by in vitro kinase assay as well as by an organ bath assay. In vivo, S-guanylation of PKG was demonstrated to occur in mice without any specific treatment and was significantly enhanced by lipopolysaccharide administration. These findings warrant further investigation in terms of the physiological and pathophysiological roles of S-guanylation-dependent persistent PKG activation.
authors.affiliation: Department of Environmental Health Sciences and Molecular Toxicology, Tohoku University Graduate School of Medicine , 2-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan.
authors.forename: Soichiro
authors.initials: S
authors.lastname: Akashi
authors.affiliation: Department of Microbiology, Graduate School of Medical Sciences, Kumamoto University , 1-1-1 Honjo, Chuo-ku, Kumamoto 860-8556, Japan.
authors.forename: Khandaker Ahtesham
authors.initials: KA
authors.lastname: Ahmed
authors.affiliation: Department of Microbiology, Graduate School of Medical Sciences, Kumamoto University , 1-1-1 Honjo, Chuo-ku, Kumamoto 860-8556, Japan.|Precursory Research for Embryonic Science and Technology, Japan Science and Technology Agency , Kawaguchi, Saitama 332-0012, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Sawa
authors.affiliation: Department of Microbiology, Graduate School of Medical Sciences, Kumamoto University , 1-1-1 Honjo, Chuo-ku, Kumamoto 860-8556, Japan.
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Ono
authors.affiliation: Department of Microbiology, Graduate School of Medical Sciences, Kumamoto University , 1-1-1 Honjo, Chuo-ku, Kumamoto 860-8556, Japan.
authors.forename: Hiroyasu
authors.initials: H
authors.lastname: Tsutsuki
authors.affiliation: Department of Cardiology, Cardiovascular Division, King's College London, The Rayne Institute, St Thomas' Hospital , London SE1 7EH, U.K.
authors.forename: Joseph R
authors.initials: JR
authors.lastname: Burgoyne
authors.affiliation: Department of Environmental Health Sciences and Molecular Toxicology, Tohoku University Graduate School of Medicine , 2-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan.
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Ida
authors.affiliation: Department of Molecular Genetics, Graduate School of Medical Sciences, Kumamoto University , Honjo 1-1-1, Kumamoto 860-8556, Japan.
authors.forename: Eiji
authors.initials: E
authors.lastname: Horio
authors.affiliation: Department of Cardiology, Cardiovascular Division, King's College London, The Rayne Institute, St Thomas' Hospital , London SE1 7EH, U.K.
authors.forename: Oleksandra
authors.initials: O
authors.lastname: Prysyazhna
authors.affiliation: Department of Molecular Genetics, Graduate School of Medical Sciences, Kumamoto University , Honjo 1-1-1, Kumamoto 860-8556, Japan.
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Oike
authors.affiliation: Department of Environmental Health Sciences and Molecular Toxicology, Tohoku University Graduate School of Medicine , 2-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan.
authors.forename: Mizanur Md
authors.initials: MM
authors.lastname: Rahaman
authors.affiliation: Department of Cardiology, Cardiovascular Division, King's College London, The Rayne Institute, St Thomas' Hospital , London SE1 7EH, U.K.
authors.forename: Philip
authors.initials: P
authors.lastname: Eaton
authors.affiliation: Department of Environmental Health Sciences and Molecular Toxicology, Tohoku University Graduate School of Medicine , 2-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan.
authors.forename: Shigemoto
authors.initials: S
authors.lastname: Fujii
authors.affiliation: Department of Environmental Health Sciences and Molecular Toxicology, Tohoku University Graduate School of Medicine , 2-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan.
authors.forename: Takaaki
authors.initials: T
authors.lastname: Akaike
mesh_terms: D000818:Animals; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism*; D004789:Enzyme Activation; D006147:Guanine / Q000378:metabolism*; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009206:Myocardium / Q000201:enzymology / Q000378:metabolism; D009712:Nucleotides, Cyclic / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*
chemical_list: D009712:Nucleotides, Cyclic; D011506:Proteins; D006147:Guanine; D017869:Cyclic GMP-Dependent Protein Kinases