The specific aims of this proposal are to obtain high-resolution structures of the cGMP- binding domains of cGMP-dependent protein kinase I and II (PKGs) and to use the structural information to design activators specific for PKGs. As key receptors for cGMP, PKG I and II mediate most effects of cGMP-elevating drugs such as nitric oxide-releasing agents for the treatment of many hypertensive diseases and phosphodiesterase inhibitors for the treatment of erectile dysfunction. While PKG I and II are therapeutic targets fo treating hypertensive diseases such as arterial and pulmonary hypertension, heart failure, erectile dysfunction and osteoporosis, developing specific activators has been difficult mainly due to a lack of available structural information. To obtain structural information needed for developing specific activators of PKG I and II, our group recently determined crystal structures of a fragment of the regulatory domains of human PKG I and II that specifically bind cGMP and activate catalytic activity. My long-term goals are to understand the activation mechanism of PKG mediated by cGMP and to develop specific activators of PKG that can be used for treating hypertensive diseases. To achieve these goals, my plans are to understand the role of the non-selective A-domain in activation of PKG I, to understand the cyclic nucleotide selectivity mechanism of PKG II, to screen known cGMP analogs for isoform specific binding and activation, and determine their co-crystal structures and use the resulting structural information for developing isoform specific activators of PKG I and II.

Public Health Relevance

cGMP-dependent protein kinase I and II (PKG) are the central mediators of NO- cGMP signaling pathway that regulates platelet aggregation, smooth muscle tone, bone growth, renin secretion and memory formation. Although PKGs can be targeted for treating diseases such as, erectile dysfunction, cardiovascular and pulmonary diseases, and osteoporosis, developing isotype specific activators and inhibitors has been difficult because there is no structural information available. My ultimate goal is to rationally target these kinase by obtaining their crystal structures in high-resolution and develop pharmacological agents that can modulate the activity of the kinase to treat diseases related to NO-cGMP signaling dysfunction.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM090161-07
Application #
9102231
Study Section
Molecular and Integrative Signal Transduction Study Section (MIST)
Program Officer
Gerratana, Barbara
Project Start
2010-07-01
Project End
2019-06-30
Budget Start
2016-07-01
Budget End
2017-06-30
Support Year
7
Fiscal Year
2016
Total Cost
Indirect Cost
Name
Baylor College of Medicine
Department
Pharmacology
Type
Schools of Medicine
DUNS #
051113330
City
Houston
State
TX
Country
United States
Zip Code
77030
Gerlits, Oksana; Campbell, James C; Blakeley, Matthew P et al. (2018) Neutron Crystallography Detects Differences in Protein Dynamics: Structure of the PKG II Cyclic Nucleotide Binding Domain in Complex with an Activator. Biochemistry 57:1833-1837
Qin, Liying; Sankaran, Banumathi; Aminzai, Sahar et al. (2018) Structural basis for selective inhibition of human PKG I? by the balanol-like compound N46. J Biol Chem 293:10985-10992
Campbell, James C; VanSchouwen, Bryan; Lorenz, Robin et al. (2017) Crystal structure of cGMP-dependent protein kinase I? cyclic nucleotide-binding-B domain : Rp-cGMPS complex reveals an apo-like, inactive conformation. FEBS Lett 591:221-230
Campbell, James C; Henning, Philipp; Franz, Eugen et al. (2017) Structural Basis of Analog Specificity in PKG I and II. ACS Chem Biol 12:2388-2398
Lorenz, Robin; Moon, Eui-Whan; Kim, Jeong Joo et al. (2017) Mutations of PKA cyclic nucleotide-binding domains reveal novel aspects of cyclic nucleotide selectivity. Biochem J 474:2389-2403
He, Daniel; Lorenz, Robin; Kim, Choel et al. (2017) Switching Cyclic Nucleotide-Selective Activation of Cyclic Adenosine Monophosphate-Dependent Protein Kinase Holoenzyme Reveals Distinct Roles of Tandem Cyclic Nucleotide-Binding Domains. ACS Chem Biol 12:3057-3066
Campbell, James C; Kim, Jeong Joo; Li, Kevin Y et al. (2016) Structural Basis of Cyclic Nucleotide Selectivity in cGMP-dependent Protein Kinase II. J Biol Chem 291:5623-33
Kim, Jeong Joo; Lorenz, Robin; Arold, Stefan T et al. (2016) Crystal Structure of PKG I:cGMP Complex Reveals a cGMP-Mediated Dimeric Interface that Facilitates cGMP-Induced Activation. Structure 24:710-720
Kim, Jeong Joo; Flueck, Christian; Franz, Eugen et al. (2015) Crystal structures of the carboxyl cGMP binding domain of the Plasmodium falciparum cGMP-dependent protein kinase reveal a novel capping triad crucial for merozoite egress. PLoS Pathog 11:e1004639
VanSchouwen, Bryan; Selvaratnam, Rajeevan; Giri, Rajanish et al. (2015) Mechanism of cAMP Partial Agonism in Protein Kinase G (PKG). J Biol Chem 290:28631-41

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