Skip to main content
OSTI.GOVU.S. Department of Energy Office of Scientific and Technical Information
U.S. Department of Energy
Office of Scientific and Technical Information
 

Sensing Domain Dynamics in Protein Kinase A-Iα Complexes by Solution X-ray Scattering

Journal Article · · Journal of Biological Chemistry
 [1];  [2];  [3];  [4]
  1. Univ. of California, San Diego, La Jolla, CA (United States). Dept. of Chemistry and Biochemistry; DOE/OSTI
  2. Univ. of California, San Diego, La Jolla, CA (United States). Dept. of Chemistry and Biochemistry
  3. Univ. of California, San Diego, La Jolla, CA (United States). Dept. of Chemistry and Biochemistry; Univ. of California, San Diego, La Jolla, CA (United States). The Howard Hughes Medical Inst. Dept. of Pharmacology
  4. Univ. of Utah, Salt Lake City, UT (United States). Dept. of Pharmacology and Toxicology
+ Show Author Affiliations
The catalytic (C) and regulatory (R) subunits of protein kinase A are exceptionally dynamic proteins. Interactions between the R- and C-subunits are regulated by cAMP binding to the two cyclic nucleotide-binding domains in the R-subunit. Mammalian cells express four different isoforms of the R-subunit (RIα, RIβ, RIIα, and RIIβ) that all interact with the C-subunit in different ways. Here, we investigate the dynamic behavior of protein complexes between RIα and C-subunits using small angle x-ray scattering. We show that a single point mutation in RIα, R333K (which alters the cAMP-binding properties of Domain B) results in a compact shape compared with the extended shape of the wild-type R∙C complex. A double mutant complex that disrupts the interaction site between the C-subunit and Domain B in RIα, RIαABR333K∙C(K285P), results in a broader P(r) curve that more closely resembles the P(r) profiles of wild-type complexes. These results together suggest that interactions between RIα Domain B and the C-subunit in the RIα∙C complex involve large scale dynamics that can be disrupted by single point mutations in both proteins. In contrast to RIα∙C complexes. Domain B in the RIIβ∙C heterodimer is not dynamic and is critical for both inhibition and complex formation. Our study highlights the functional differences of domain dynamics between protein kinase A isoforms, providing a framework for elucidating the global organization of each holoenzyme and the cross-talk between the R- and C-subunits.
Research Organization:
Univ. of Utah, Salt Lake City, UT (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division
Grant/Contract Number:
FG02-05ER64026
OSTI ID:
1625067
Journal Information:
Journal of Biological Chemistry, Journal Name: Journal of Biological Chemistry Journal Issue: 51 Vol. 284; ISSN 0021-9258
Publisher:
American Society for Biochemistry and Molecular BiologyCopyright Statement
Country of Publication:
United States
Language:
English

References (41)

The T cell enigma in lupus journal January 1996
Hotspots for spontaneous and mutagen-induced lesions in regulatory subunit of cyclic AMP-dependent protein kinase in S49 mouse lymphoma cells journal November 1985
Conformational Differences Among Solution Structures of the Type Iα, IIα and IIβ Protein Kinase A Regulatory Subunit Homodimers: Role of the Linker Regions journal April 2004
Mapping Intersubunit Interactions of the Regulatory Subunit (RIα) in the Type I Holoenzyme of Protein Kinase A by Amide Hydrogen/Deuterium Exchange Mass Spectrometry (DXMS) journal July 2004
Solution Scattering Reveals Large Differences in the Global Structures of Type II Protein Kinase A Isoforms journal March 2006
R-subunit Isoform Specificity in Protein Kinase A: Distinct Features of Protein Interfaces in PKA Types I and II by Amide H/2H Exchange Mass Spectrometry journal November 2007
Dynamic Binding of PKA Regulatory Subunit RIα journal January 2006
Restoring Low Resolution Structure of Biological Macromolecules from Solution Scattering Using Simulated Annealing journal June 1999
The hallmark of AGC kinase functional divergence is its C-terminal tail, a cis-acting regulatory module journal January 2007
Contribution of Non-catalytic Core Residues to Activity and Regulation in Protein Kinase A journal January 2009
Expression of the catalytic subunit of cAMP-dependent protein kinase in Escherichia coli : multiple isozymes reflect different phosphorylation states journal January 1993
The Essential Role of RIα in the Maintenance of Regulated PKA Activity journal June 2002
Phosphorylation-Dephosphorylation of Enzymes journal June 1979
Large Deletions of the PRKAR1A Gene in Carney Complex journal January 2008
Deficient type I protein kinase A isozyme activity in systemic lupus erythematosus T lymphocytes. journal July 1994
A PRKAR1A Mutation Associated with Primary Pigmented Nodular Adrenocortical Disease in 12 Kindreds journal May 2006
Identifying Critical Non-Catalytic Residues that Modulate Protein Kinase A Activity journal March 2009
Hotspots for spontaneous and mutagen-induced lesions in regulatory subunit of cyclic AMP-dependent protein kinase in S49 mouse lymphoma cells journal November 1985
Restoring Low Resolution Structure of Biological Macromolecules from Solution Scattering Using Simulated Annealing journal June 1999
Activation of type I cyclic AMP-dependent protein kinases with defective cyclic AMP-binding sites. journal February 1987
PKA-I Holoenzyme Structure Reveals a Mechanism for cAMP-Dependent Activation journal September 2007
Protein kinase A as a therapeutic target for memory disorders: rationale and challenges journal March 2005
RIα Subunit of PKA journal June 2004
The Solution Structure of a Cardiac Troponin C−Troponin I−Troponin T Complex Shows a Somewhat Compact Troponin C Interacting with an Extended Troponin I−Troponin T Component † journal December 2002
Active Site Mutations Define the Pathway for the Cooperative Activation of cAMP-Dependent Protein Kinase † journal January 1996
Compensatory Regulation of RIα Protein Levels in Protein Kinase A Mutant Mice journal February 1997
Dissecting cAMP Binding Domain A in the RIα Subunit of cAMP-dependent Protein Kinase journal October 1998
The Complete Pathway for Catalytic Activation of the Mitogen-activated Protein Kinase, ERK2 journal November 2001
cAMP-dependent Protein Kinase Regulatory Subunit Type IIβ journal February 2004
The Conformationally Dynamic C Helix of the RIα Subunit of Protein Kinase A Mediates Isoform-specific Domain Reorganization upon C Subunit Binding journal October 2005
Contribution of Non-catalytic Core Residues to Activity and Regulation in Protein Kinase A journal January 2009
Automated matching of high- and low-resolution structural models journal February 2001
Uniqueness of ab initio shape determination in small-angle scattering journal April 2003
PRIMUS: a Windows PC-based system for small-angle scattering data analysis journal September 2003
DAMMIF, a program for rapid ab-initio shape determination in small-angle scattering journal January 2009
Determination of the regularization parameter in indirect-transform methods using perceptual criteria journal August 1992
CRYSOL – a Program to Evaluate X-ray Solution Scattering of Biological Macromolecules from Atomic Coordinates journal December 1995
Crystal Structure of a Complex Between the Catalytic and Regulatory (RI ) Subunits of PKA journal February 2005
Large Deletions of the PRKAR1A Gene in Carney Complex journal January 2008
Deficient type I protein kinase A isozyme activity in systemic lupus erythematosus T lymphocytes. journal July 1994
Mutations in Regulatory Subunit Type 1A of Cyclic Adenosine 5′-Monophosphate-Dependent Protein Kinase ( PRKAR1A ): Phenotype Analysis in 353 Patients and 80 Different Genotypes journal June 2009

Cited By (1)

Switching of the folding-energy landscape governs the allosteric activation of protein kinase A journal July 2018

Similar Records

The roles of RIIbeta linker and N-terminal cyclic nucleotide-binding domain in determining the unique structures of Type IIbeta Protein Kinase A. A small angle X-ray and neutron scattering study
Journal Article · Sun Aug 10 20:00:00 EDT 2014 · Journal of Biological Chemistry · OSTI ID:1163593
Structural analyses of the PKA RIIβ holoenzyme containing the oncogenic DnaJB1-PKAc fusion protein reveal protomer asymmetry and fusion-induced allosteric perturbations in fibrolamellar hepatocellular carcinoma
Journal Article · Sun Dec 27 19:00:00 EST 2020 · PLoS Biology (Online) · OSTI ID:1903844
Tyrosine-371 contributes to the positive cooperativity between the two cAMP binding sites in the regulatory subunit of cAMP-dependent protein kinase I
Journal Article · Mon Mar 07 23:00:00 EST 1988 · Biochemistry; (United States) · OSTI ID:5317459

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
59 BASIC BIOLOGICAL SCIENCES
Biochemistry & Molecular Biology