1 research outputs found
Persistent Activation of cGMP-Dependent Protein Kinase by a Nitrated Cyclic Nucleotide via Site Specific Protein <i>S</i>‑Guanylation
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 <i>S</i>-guanylation). Here, we demonstrate that 8-nitro-cGMP potentially <i>S</i>-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. <i>S</i>-Guanylation of PKG was found to occur in a site specific
manner; Cys42 and Cys195 were the susceptible residues among 11 Cys
residues. Importantly, <i>S</i>-guanylation at Cys195, which
is located in the high-affinity cGMP binding domain of PKG, causes
persistent enzyme activation as determined by <i>in vitro</i> kinase assay as well as by an organ bath assay. <i>In vivo</i>, <i>S</i>-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 <i>S</i>-guanylation-dependent persistent PKG activation