Neutron Diffraction Reveals Hydrogen Bonds Critical for cGMP-Selective Activation: Insights for cGMP-Dependent Protein Kinase Agonist Design

Huang, Gilbert Y.; Gerlits, Oksana O.; Blakeley, Matthew P.; Sankaran, Banumathi; Kovalevsky, Andrey Y.; Kim, Choel

doi:10.1021/bi501012v

Title: Neutron Diffraction Reveals Hydrogen Bonds Critical for cGMP-Selective Activation: Insights for cGMP-Dependent Protein Kinase Agonist Design

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Abstract

High selectivity of cyclic-nucleotide binding (CNB) domains for cAMP and cGMP are required for segregating signaling pathways; however, the mechanism of selectivity remains unclear. To investigate the mechanism of high selectivity in cGMP-dependent protein kinase (PKG), we determined a room-temperature joint X-ray/neutron (XN) structure of PKG Iβ CNB-B, a domain 200-fold selective for cGMP over cAMP, bound to cGMP (2.2 Å), and a low-temperature X-ray structure of CNB-B with cAMP (1.3 Å). Finally, the XN structure directly describes the hydrogen bonding interactions that modulate high selectivity for cGMP, while the structure with cAMP reveals that all these contacts are disrupted, explaining its low affinity for cAMP.

Authors:

Huang, Gilbert Y. ^[1]; Gerlits, Oksana O. ^[2]; Blakeley, Matthew P. ^[3]; Sankaran, Banumathi ^[4]; Kovalevsky, Andrey Y. ^[2]; Kim, Choel ^[5]

Verna and Mars McClean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, Texas 77004, United States
Biology and Soft Matter Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831, United States
Institute Laue-Langevin, 71 Avenue des Martyrs, 38000, Grenoble, France
Berkeley Center for Structural Biology, Lawrence Berkeley National Laboratory, Berkeley, California, United States
Verna and Mars McClean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, Texas 77004, United States, Department of Pharmacology, Baylor College of Medicine, One Baylor Plaza, Houston, Texas 77004, United States

Publication Date:: 2014-10-22

Research Org.:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Org.:: USDOE Office of Science (SC), Basic Energy Sciences (BES)

OSTI Identifier:: 1163250

Alternate Identifier(s):: OSTI ID: 1257634

Grant/Contract Number:: AC02-05CH11231; R01GM90161

Resource Type:: Published Article

Journal Name:: Biochemistry

Additional Journal Information:: Journal Name: Biochemistry Journal Volume: 53 Journal Issue: 43; Journal ID: ISSN 0006-2960

Publisher:: American Chemical Society

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES

Citation Formats


                    Huang, Gilbert Y., Gerlits, Oksana O., Blakeley, Matthew P., Sankaran, Banumathi, Kovalevsky, Andrey Y., and Kim, Choel. Neutron Diffraction Reveals Hydrogen Bonds Critical for cGMP-Selective Activation: Insights for cGMP-Dependent Protein Kinase Agonist Design.  United States: N. p., 2014. 
Web.  doi:10.1021/bi501012v.

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                    Huang, Gilbert Y., Gerlits, Oksana O., Blakeley, Matthew P., Sankaran, Banumathi, Kovalevsky, Andrey Y., & Kim, Choel. Neutron Diffraction Reveals Hydrogen Bonds Critical for cGMP-Selective Activation: Insights for cGMP-Dependent Protein Kinase Agonist Design.  United States.  https://doi.org/10.1021/bi501012v

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                    Huang, Gilbert Y., Gerlits, Oksana O., Blakeley, Matthew P., Sankaran, Banumathi, Kovalevsky, Andrey Y., and Kim, Choel. Wed .  
"Neutron Diffraction Reveals Hydrogen Bonds Critical for cGMP-Selective Activation: Insights for cGMP-Dependent Protein Kinase Agonist Design".  United States.  https://doi.org/10.1021/bi501012v.

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@article{osti_1163250,

  title        = {Neutron Diffraction Reveals Hydrogen Bonds Critical for cGMP-Selective Activation: Insights for cGMP-Dependent Protein Kinase Agonist Design},

  author       = {Huang, Gilbert Y. and Gerlits, Oksana O. and Blakeley, Matthew P. and Sankaran, Banumathi and Kovalevsky, Andrey Y. and Kim, Choel},

  abstractNote = {High selectivity of cyclic-nucleotide binding (CNB) domains for cAMP and cGMP are required for segregating signaling pathways; however, the mechanism of selectivity remains unclear. To investigate the mechanism of high selectivity in cGMP-dependent protein kinase (PKG), we determined a room-temperature joint X-ray/neutron (XN) structure of PKG Iβ CNB-B, a domain 200-fold selective for cGMP over cAMP, bound to cGMP (2.2 Å), and a low-temperature X-ray structure of CNB-B with cAMP (1.3 Å). Finally, the XN structure directly describes the hydrogen bonding interactions that modulate high selectivity for cGMP, while the structure with cAMP reveals that all these contacts are disrupted, explaining its low affinity for cAMP.},

  doi          = {10.1021/bi501012v},

  journal      = {Biochemistry},

  number       = 43,

  volume       = 53,

  place        = {United States},

  year         = {2014},

  month        = {10}

}

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