An N-terminally truncated form of cyclic GMP–dependent protein kinase Iα (PKG Iα) is monomeric and autoinhibited and provides a model for activation
AuthorMoon, Thomas M.
Sheehe, Jessica L.
Nausch, Lydia W.
Matthews, Dwight E.
Blumenthal, Donald K.
Dostmann, Wolfgang R.
AffiliationUniv Arizona, Dept Chem & Biochem
Keywordscyclic GMP (cGMP)
small-angle X-ray scattering (SAXS)
surface plasmon resonance (SPR)
cGMP-dependent protein kinase
MetadataShow full item record
CitationMoon, T. M., Sheehe, J. L., Nukareddy, P., Nausch, L. W., Wohlfahrt, J., Matthews, D. E., ... & Dostmann, W. R. (2018). An N-terminally truncated form of cyclic GMP-dependent protein kinase Iα (PKG Iα) is monomeric, autoinhibited, and provides a model for activation. J. Biol. Chem. 2018 293: 7916-7929. doi:10.1074/jbc.RA117.000647
JournalJOURNAL OF BIOLOGICAL CHEMISTRY
Rights© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.
Collection InformationThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at firstname.lastname@example.org.
AbstractThe type I cGMP-dependent protein kinases (PKG I) serve essential physiological functions, including smooth muscle relaxation, cardiac remodeling, and platelet aggregation. These enzymes form homodimers through their N-terminal dimerization domains, a feature implicated in regulating their cooperative activation. Previous investigations into the activation mechanisms of PKG I isoforms have been largely influenced by structures of the cAMP-dependent protein kinase (PKA). Here, we examined PKG I activation by cGMP and cAMP by engineering a monomeric form that lacks N-terminal residues 1-53 (53). We found that the construct exists as a monomer as assessed by whole-protein MS, size-exclusion chromatography, and small-angle X-ray scattering (SAXS). Reconstruction of the SAXS 3D envelope indicates that 53 has a similar shape to the heterodimeric RI-C complex of PKA. Moreover, we found that the 53 construct is autoinhibited in its cGMP-free state and can bind to and be activated by cGMP in a manner similar to full-length PKG I as assessed by surface plasmon resonance (SPR) spectroscopy. However, we found that the 53 variant does not exhibit cooperative activation, and its cyclic nucleotide selectivity is diminished. These findings support a model in which, despite structural similarities, PKG I activation is distinct from that of PKA, and its cooperativity is driven by in trans interactions between protomers.
Note12 month embargo; published online: 30 March 2018
VersionFinal published version
SponsorsUnited States Department of Energy (DOE), Office of Science, Office of Basic Energy Sciences [DE-AC02-76SF00515]; DOE Office of Biological and Environmental Research; National Institute of General Medical Sciences at the National Institutes of Health (NIH) [P41GM103393]; Centers of Biomedical Research Excellence (COBRE) from the NIGMS, National Institutes of Health [P30-GM118228]
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