skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Co-Crystal Structures of PKG Iβ (92–227) with cGMP and cAMP Reveal the Molecular Details of Cyclic-Nucleotide Binding

Journal Article · · PLoS ONE
 [1];  [2];  [3];  [1];  [4];  [5];  [6];  [3];  [3];  [7];  [1]
  1. Baylor Univ., Houston, TX (United States). College of Medicine. Dept. of Pharmacology
  2. Univ. of California, San Diego, La Jolla, CA (United States). Dept. of Medicine
  3. Baylor Univ., Houston, TX (United States). College of Medicine. The Verna and Marrs McLean Dept. of Biochemistry and Molecular Biology
  4. Rice Univ., Houston, TX (United States)
  5. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). The Berkeley Center for Structural Biology
  6. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Computational Crystallography Initiative
  7. Baylor Univ., Houston, TX (United States). College of Medicine. Dept. of Pharmacology; Baylor Univ., Houston, TX (United States). College of Medicine. The Verna and Marrs McLean Dept. of Biochemistry and Molecular Biology; Baylor Univ., Houston, TX (United States). College of Medicine. Dept. of Molecular Virology and Microbiology
+ Show Author Affiliations

Background: Cyclic GMP-dependent protein kinases (PKGs) are central mediators of the NO-cGMP signaling pathway and phosphorylate downstream substrates that are crucial for regulating smooth muscle tone, platelet activation, nociception and memory formation. As one of the main receptors for cGMP, PKGs mediate most of the effects of cGMP elevating drugs, such as nitric oxide-releasing agents and phosphodiesterase inhibitors which are used for the treatment of angina pectoris and erectile dysfunction, respectively. Methodology/Principal Findings: We have investigated the mechanism of cyclic nucleotide binding to PKG by determining crystal structures of the amino-terminal cyclic nucleotide-binding domain (CNBD-A) of human PKG I bound to either cGMP or cAMP. We also determined the structure of CNBD-A in the absence of bound nucleotide. The crystal structures of CNBD-A with bound cAMP or cGMP reveal that cAMP binds in either syn or anti configurations whereas cGMP binds only in a syn configuration, with a conserved threonine residue anchoring both cyclic phosphate and guanine moieties. The structure of CNBD-A in the absence of bound cyclic nucleotide was similar to that of the cyclic nucleotide bound structures. Surprisingly, isothermal titration calorimetry experiments demonstrated that CNBD-A binds both cGMP and cAMP with a relatively high affinity, showing an approximately two-fold preference for cGMP. Conclusions/Significance: Our findings suggest that CNBD-A binds cGMP in the syn conformation through its interaction with Thr193 and an unusual cis-peptide forming residues Leu172 and Cys173. Although these studies provide the first structural insights into cyclic nucleotide binding to PKG, our ITC results show only a two-fold preference for cGMP, indicating that other domains are required for the previously reported cyclic nucleotide selectivity.

Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division; National Institutes of Health (NIH)
Grant/Contract Number:
AC02-05CH11231; K22-CA124517
OSTI ID:
1629608
Journal Information:
PLoS ONE, Vol. 6, Issue 4; ISSN 1932-6203
Publisher:
Public Library of ScienceCopyright Statement
Country of Publication:
United States
Language:
English

References (42)

Crystal structure of a CAP-DNA complex: the DNA is bent by 90 degrees journal August 1991
Activation by Cyclic GMP Binding Causes an Apparent Conformational Change in cGMP-dependent Protein Kinase journal December 1997
A simple electrostatic switch important in the activation of type I protein kinase A by cyclic AMP journal January 2006
Fast and Slow Cyclic Nucleotide-dissociation Sites in cAMP-dependent Protein Kinase Are Transposed in Type Iβ cGMP-dependent Protein Kinase journal July 1996
PHENIX: a comprehensive Python-based system for macromolecular structure solution. text January 2010
Cavin1 intrinsically disordered domains are essential for fuzzy electrostatic interactions and caveola formation journal February 2021
cGMP-binding Prepares PKG for Substrate Binding by Disclosing the C-terminal Domain journal February 2008
MolProbity : all-atom structure validation for macromolecular crystallography journal December 2009
Cyclic Nucleotide-Dependent Protein Kinases: Intracellular Receptors for cAMP and cGMP Action journal January 1999
Structural genomics of human proteins – target selection and generation of a public catalogue of expression clones journal July 2005
Coot model-building tools for molecular graphics journal November 2004
Use of TLS parameters to model anisotropic displacements in macromolecular refinement journal January 2001
Isolated Regulatory Domains of cGMP-dependent Protein Kinase Iα and Iβ Retain Dimerization and Native cGMP-binding Properties and Undergo Isoform-specific Conformational Changes journal January 2006
The activation of expressed cGMP-dependent protein kinase isozymes Ialpha and Ibeta is determined by the different amino-termini journal December 1991
Exploring the gas access routes in a [NiFeSe] hydrogenase using crystals pressurized with krypton and oxygen journal August 2020
Predicted structures of the cGMP binding domains of the cGMP-dependent protein kinase: a key alanine/threonine difference in evolutionary divergence of cAMP and cGMP binding sites journal July 1989
PHENIX: a comprehensive Python-based system for macromolecular structure solution journal January 2010
Function of cGMP-Dependent Protein Kinases as Revealed by Gene Deletion journal January 2006
Determination of the syn-anti Equilibrium of Some Purine 3':5'-Nucleotides by Nuclear-Magnetic-Relaxation Perturbation in the Presence of a Lanthanide-Ion Probe journal June 1977
cGMP-dependent protein kinase. Autophosphorylation changes the characteristics of binding site 1 journal March 1985
Iterative-build OMIT maps: map improvement by iterative model building and refinement without model bias. text January 2008
Identification of the Amino Acid Sequences Responsible for High Affinity Activation of cGMP Kinase Iα journal April 1997
PKA-I Holoenzyme Structure Reveals a Mechanism for cAMP-Dependent Activation journal September 2007
Mechanisms associated with cGMP binding and activation of cGMP-dependent protein kinase journal February 2003
Iterative-build OMIT maps: map improvement by iterative model building and refinement without model bias journal April 2008
Regulatory subunit of protein kinase A: structure of deletion mutant with cAMP binding domains journal August 1995
Distinguishing the Roles of the Two Different cGMP-binding Sites for Modulating Phosphorylation of Exogenous Substrate (Heterophosphorylation) and Autophosphorylation of cGMP-dependent Protein Kinase journal January 2000
Crystal Structure of a Complex Between the Catalytic and Regulatory (RI ) Subunits of PKA journal February 2005
Control of ligand specificity in cyclic nucleotide-gated channels from rod photoreceptors and olfactory epithelium. journal November 1991
Structural basis for modulation and agonist specificity of HCN pacemaker channels journal September 2003
Automated Structure Solution with the PHENIX Suite book January 2008
Super-resolution biomolecular crystallography with low-resolution data journal April 2010
Rising behind NO: cGMP-dependent protein kinases journal May 2000
Studies of cGMP analog specificity and function of the two intrasubunit binding sites of cGMP-dependent protein kinase. journal January 1986
Mutating protein kinase cAMP-binding sites into cGMP-binding sites. Mechanism of cGMP selectivity. journal December 1991
Oxidation of cysteines activates cGMP-dependent protein kinase. journal September 1991
Characterization of a novel isozyme of cGMP-dependent protein kinase from bovine aorta journal May 1989
Properties of a cGMP-dependent Monomeric Protein Kinase from Bovine Aorta journal March 1989
From Poor Resolution to Rich Insight journal June 2010
Conformations of cyclic 3′,5′-nucleotides. Effect of the base on the syn-anti conformer distribution journal January 1974
A Conserved Serine Juxtaposed to the Pseudosubstrate Site of Type I cGMP-dependent Protein Kinase Contributes Strongly to Autoinhibition and Lower cGMP Affinity journal September 2002
Dimerization of cGMP-dependent Protein Kinase Iβ Is Mediated by an Extensive Amino-terminal Leucine Zipper Motif, and Dimerization Modulates Enzyme Function journal December 2003

Cited By (15)

Structures of the cGMP-dependent protein kinase in malaria parasites reveal a unique structural relay mechanism for activation journal June 2019
Nucleotide signaling pathway convergence in a cAMP‐sensing bacterial c‐di‐GMP phosphodiesterase journal July 2019
An N-terminally truncated form of cyclic GMP–dependent protein kinase Iα (PKG Iα) is monomeric and autoinhibited and provides a model for activation journal March 2018
Phosphodiesterase 1 Bridges Glutamate Inputs with NO- and Dopamine-Induced Cyclic Nucleotide Signals in the Striatum journal March 2019
Correction to: Crystal structure of PKG Iβ holoenzyme reveals a trans‑inhibiting dimer assembly journal January 2020
A Mechanism for the Auto-inhibition of Hyperpolarization-activated Cyclic Nucleotide-gated (HCN) Channel Opening and Its Relief by cAMP journal August 2014
cAMP-Dependent Protein Kinase and cGMP-Dependent Protein Kinase as Cyclic Nucleotide Effectors book January 2015
Structures of the cGMP-dependent protein kinase in malaria parasites reveal a unique structural relay mechanism for activation. text January 2020
A Secondary Structural Transition in the C-helix Promotes Gating of Cyclic Nucleotide-regulated Ion Channels journal March 2013
The role of cGMP signalling in regulating life cycle progression of Plasmodium journal August 2012
Neutron Diffraction Reveals Hydrogen Bonds Critical for cGMP-Selective Activation: Insights for cGMP-Dependent Protein Kinase Agonist Design journal October 2014
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 journal February 2015
Establishing a Split Luciferase Assay for Proteinkinase G (PKG) Interaction Studies journal April 2018
Funktion von IRAG für cGMP-Kinase-I-Komplexe sowie für die InsP3R-I-Phosphorylierung text January 2012
Identification of cGMP-Kinase Complexes by Affinity Chromatography book March 2012

Similar Records

Related Subjects

59 BASIC BIOLOGICAL SCIENCES