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Title: Crystal structure of cGMP-dependent protein kinase Iβ cyclic nucleotide-binding-B domain : Rp-cGMPS complex reveals an apo-like, inactive conformation

The R-diastereomer of phosphorothioate analogs of cGMP, Rp-cGMPS, is one of few known inhibitors of cGMP-dependent protein kinase I (PKG I); however, its mechanism of inhibition is currently not fully understood. We determined the crystal structure of the PKG Iβ cyclic nucleotide-binding domain (PKG Iβ CNB-B), considered a ‘gatekeeper’ for cGMP activation, bound to Rp-cGMPS at 1.3 Å. Our structural and NMR data show that PKG Iβ CNB-B bound to Rp-cGMPS displays an apo-like structure with its helical domain in an open conformation. Comparison with the cAMP-dependent protein kinase regulatory subunit (PKA RIα) showed that this conformation resembles the catalytic subunit-bound inhibited state of PKA RIα more closely than the apo or Rp-cAMPS-bound conformations. Our results suggest that Rp-cGMPS inhibits PKG I by stabilizing the inactive conformation of CNB-B.
Authors:
 [1] ;  [2] ;  [3] ;  [4] ;  [3] ;  [2] ;  [5]
  1. Baylor College of Medicine, Houston, TX (United States). Structural and Computational Biology and Molecular Biophysics Program, Dept. of Chemistry and Chemical Biology
  2. McMaster Univ., Hamilton, ON (Canada). Dept. of Chemistry and Chemical Biology
  3. Univ. of Kassel, Hesse (Germany). Dept. of Biochemistry
  4. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Berkeley Center for Structural Biology
  5. Baylor College of Medicine, Houston, TX (United States). Structural and Computational Biology and Molecular Biophysics Program, Dept. of Chemistry and Chemical Biology, Verna and Marrs McLean Dept. of Biochemistry and Molecular Biology
Publication Date:
Grant/Contract Number:
AC02-05CH11231
Type:
Accepted Manuscript
Journal Name:
FEBS Letters
Additional Journal Information:
Journal Volume: 591; Journal Issue: 1; Journal ID: ISSN 0014-5793
Publisher:
Federation of European Biochemical Societies
Research Org:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); National Institutes of Health (NIH)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; cGMP dependent protein kinase; PKG; second messengers; NO-cGMP signaling; kinase inhibition
OSTI Identifier:
1415962
Alternate Identifier(s):
OSTI ID: 1399282

Campbell, James C., VanSchouwen, Bryan, Lorenz, Robin, Sankaran, Banumathi, Herberg, Friedrich W., Melacini, Giuseppe, and Kim, Choel. Crystal structure of cGMP-dependent protein kinase Iβ cyclic nucleotide-binding-B domain : Rp-cGMPS complex reveals an apo-like, inactive conformation. United States: N. p., Web. doi:10.1002/1873-3468.12505.
Campbell, James C., VanSchouwen, Bryan, Lorenz, Robin, Sankaran, Banumathi, Herberg, Friedrich W., Melacini, Giuseppe, & Kim, Choel. Crystal structure of cGMP-dependent protein kinase Iβ cyclic nucleotide-binding-B domain : Rp-cGMPS complex reveals an apo-like, inactive conformation. United States. doi:10.1002/1873-3468.12505.
Campbell, James C., VanSchouwen, Bryan, Lorenz, Robin, Sankaran, Banumathi, Herberg, Friedrich W., Melacini, Giuseppe, and Kim, Choel. 2016. "Crystal structure of cGMP-dependent protein kinase Iβ cyclic nucleotide-binding-B domain : Rp-cGMPS complex reveals an apo-like, inactive conformation". United States. doi:10.1002/1873-3468.12505. https://www.osti.gov/servlets/purl/1415962.
@article{osti_1415962,
title = {Crystal structure of cGMP-dependent protein kinase Iβ cyclic nucleotide-binding-B domain : Rp-cGMPS complex reveals an apo-like, inactive conformation},
author = {Campbell, James C. and VanSchouwen, Bryan and Lorenz, Robin and Sankaran, Banumathi and Herberg, Friedrich W. and Melacini, Giuseppe and Kim, Choel},
abstractNote = {The R-diastereomer of phosphorothioate analogs of cGMP, Rp-cGMPS, is one of few known inhibitors of cGMP-dependent protein kinase I (PKG I); however, its mechanism of inhibition is currently not fully understood. We determined the crystal structure of the PKG Iβ cyclic nucleotide-binding domain (PKG Iβ CNB-B), considered a ‘gatekeeper’ for cGMP activation, bound to Rp-cGMPS at 1.3 Å. Our structural and NMR data show that PKG Iβ CNB-B bound to Rp-cGMPS displays an apo-like structure with its helical domain in an open conformation. Comparison with the cAMP-dependent protein kinase regulatory subunit (PKA RIα) showed that this conformation resembles the catalytic subunit-bound inhibited state of PKA RIα more closely than the apo or Rp-cAMPS-bound conformations. Our results suggest that Rp-cGMPS inhibits PKG I by stabilizing the inactive conformation of CNB-B.},
doi = {10.1002/1873-3468.12505},
journal = {FEBS Letters},
number = 1,
volume = 591,
place = {United States},
year = {2016},
month = {12}
}