期刊
ELIFE
卷 11, 期 -, 页码 -出版社
eLIFE SCIENCES PUBL LTD
DOI: 10.7554/eLife.79530
关键词
cGMP signaling; kinase regulation; mammalian signal transduction; second messenger; protein-protein complex; TAAD; kinase activation; Human
类别
资金
- National Institute of General Medical Sciences [R01 GM090161]
- Deutsche Forschungsgemeinschaft [He1818/10-1]
- Federal Ministry of Education and Research, Germany
- Federal Ministry of Education and Research [FKZ: 16GW0270]
- Canadian Institutes of Health Research [389522]
This study reveals the mechanisms of auto-inhibition and cGMP relief in PKG I beta through crystal structures and biochemical analysis. The findings suggest that auto-inhibition of PKG I beta is mediated by internal contacts within each monomer, and a model for the regulation and cooperative activation of PKGs is proposed based on the available structural and biochemical data.
Cyclic GMP-dependent protein kinases (PKGs) are key mediators of the nitric oxide/cyclic guanosine monophosphate (cGMP) signaling pathway that regulates biological functions as diverse as smooth muscle contraction, cardiac function, and axon guidance. Understanding how cGMP differentially triggers mammalian PKG isoforms could lead to new therapeutics that inhibit or activate PKGs, complementing drugs that target nitric oxide synthases and cyclic nucleotide phosphodiesterases in this signaling axis. Alternate splicing of PRKG1 transcripts confers distinct leucine zippers, linkers, and auto-inhibitory (AI) pseudo-substrate sequences to PKG I alpha and I beta that result in isoform-specific activation properties, but the mechanism of enzyme auto-inhibition and its alleviation by cGMP is not well understood. Here, we present a crystal structure of PKG I beta in which the AI sequence and the cyclic nucleotide-binding (CNB) domains are bound to the catalytic domain, providing a snapshot of the auto-inhibited state. Specific contacts between the PKG I beta AI sequence and the enzyme active site help explain isoform-specific activation constants and the effects of phosphorylation in the linker. We also present a crystal structure of a PKG I CNB domain with an activating mutation linked to Thoracic Aortic Aneurysms and Dissections. Similarity of this structure to wildtype cGMP-bound domains and differences with the auto-inhibited enzyme provide a mechanistic basis for constitutive activation. We show that PKG I beta auto-inhibition is mediated by contacts within each monomer of the native full-length dimeric protein, and using the available structural and biochemical data we develop a model for the regulation and cooperative activation of PKGs.
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