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Zooming in on protons: Neutron structure of protein kinase A trapped in a product complex

Journal Article · · Science Advances
 [1];  [2];  [3];  [4];  [5];  [2]
  1. Univ. of Tennessee, Knoxville, TN (United States). Bredesen Center
  2. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Neutron Scattering Division
  3. Inst. Laue-Langevin, Grenoble (France). Large Scale Structures Group
  4. Univ. of Minnesota, Minneapolis, MN (United States). Dept. of Chemistry; Univ. of Minnesota, Minneapolis, MN (United States). Dept. of Biochemistry, Molecular Biology, and Biophysics
  5. Univ. of California, San Diego, CA (United States). Dept. of Pharmacology, and Dept. of Chemistry and Biochemistry
+ Show Author Affiliations

The question vis-à-vis the chemistry of phosphoryl group transfer catalyzed by protein kinases remains a major challenge. The neutron diffraction structure of the catalytic subunit of cAMP-dependent protein kinase (PKA-C) provides a more complete chemical portrait of key proton interactions at the active site. By using a high-affinity protein kinase substrate (PKS) peptide, we captured the reaction products, dephosphorylated nucleotide [adenosine diphosphate (ADP)] and phosphorylated PKS (pPKS), bound at the active site. In the complex, the phosphoryl group of the peptide is protonated, whereas the carboxyl group of the catalytic Asp166is not. Our structure, including conserved waters, shows how the peptide links the distal parts of the cleft together, creating a network that engages the entire molecule. By comparing slow-exchanging backbone amides to those determined by the NMR analysis of PKA-C with ADP and inhibitor peptide (PKI), we identified exchangeable amides that likely distinguish catalytic and inhibited states.

Research Organization:
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Grant/Contract Number:
AC05-00OR22725
OSTI ID:
1506775
Journal Information:
Science Advances, Journal Name: Science Advances Journal Issue: 3 Vol. 5; ISSN 2375-2548
Publisher:
AAASCopyright Statement
Country of Publication:
United States
Language:
English

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Cited By (1)

A shared vision for macromolecular crystallography over the next five years journal November 2019

Figures / Tables (6)

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37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY