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

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

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:
 [1];  [2];  [3];  [4];  [2];  [5]
  1. Verna and Mars McClean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, Texas 77004, United States
  2. Biology and Soft Matter Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831, United States
  3. Institute Laue-Langevin, 71 Avenue des Martyrs, 38000, Grenoble, France
  4. Berkeley Center for Structural Biology, Lawrence Berkeley National Laboratory, Berkeley, California, United States
  5. 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:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
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.
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. doi:10.1021/bi501012v.
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. doi:10.1021/bi501012v.
@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}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
DOI: 10.1021/bi501012v

Citation Metrics:
Cited by: 12 works
Citation information provided by
Web of Science

Save / Share:

Works referencing / citing this record:

Structures of the cGMP-dependent protein kinase in malaria parasites reveal a unique structural relay mechanism for activation
journal, June 2019

  • El Bakkouri, Majida; Kouidmi, Imène; Wernimont, Amy K.
  • Proceedings of the National Academy of Sciences, Vol. 116, Issue 28
  • DOI: 10.1073/pnas.1905558116

Structures of the cGMP-dependent protein kinase in malaria parasites reveal a unique structural relay mechanism for activation
journal, June 2019

  • El Bakkouri, Majida; Kouidmi, Imène; Wernimont, Amy K.
  • Proceedings of the National Academy of Sciences, Vol. 116, Issue 28
  • DOI: 10.1073/pnas.1905558116