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TitleMultiplicity of Regulatory Subunit Conformations Defines Structural Ensemble of Reset Protein Kinase A Holoenzyme.
Journal, issue, pagesJ Am Chem Soc, Vol. 147, Issue 17, Page 14174-14190, Year 2025
Publish dateApr 30, 2025
AuthorsVarun Venkatakrishnan / Tatiana N Laremore / Theresa S C Buckley / Jean-Paul Armache / Ganesh S Anand /
PubMed AbstractHow protein kinase A (PKA) is reset to a basal state following 3'5'-cyclic adenosine monophosphate (cAMP)-mediated activation is unknown. Here we describe the mechanism of cAMP-PKA type I signal ...How protein kinase A (PKA) is reset to a basal state following 3'5'-cyclic adenosine monophosphate (cAMP)-mediated activation is unknown. Here we describe the mechanism of cAMP-PKA type I signal termination leading to a reset of PKA by holoenzyme formation through the obligatory action of phosphodiesterases (PDEs). We report a catalytic subunit (Cα)-assisted mechanism for the reset of type I PKA and describe for the first time multiple structures of the reset PKA holoenzyme (RIα:Cα) that capture an ensemble of multiple conformational end-states through integrative electron microscopy and structural mass spectrometry approaches. Together these complementary methods highlight the large conformational dynamics of the regulatory subunit (RIα) within the tetrameric reset PKA holoenzyme. The cAMP-free reset PKA holoenzyme adopts multiple distinct conformations of RIα with contributions from the N-terminal linker and CNB-B dynamics. Our findings highlight the interplay between RIα, Cα, and PDEs (PDE8) in cAMP-PKA signalosomes to offer a new paradigm for PDE-mediated regulation of cAMP-PKA signaling.
External linksJ Am Chem Soc / PubMed:40241376
MethodsEM (single particle)
Resolution4.18 - 6.3 Å
Structure data

47944

9edc

EMDB-47944, PDB-9edc:
Reset Type-I Protein Kinase A Holoenzyme
Method: EM (single particle) / Resolution: 4.18 Å

47945

9edd

EMDB-47945, PDB-9edd:
Reset Type-I Protein Kinase A Holoenzyme
Method: EM (single particle) / Resolution: 6.3 Å

47946

9ede

EMDB-47946, PDB-9ede:
Reset Type-I Protein Kinase A Holoenzyme
Method: EM (single particle) / Resolution: 5.9 Å

Chemicals

ChemComp-ATP:
ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying molecule*YM

Source
  • homo sapiens (human)
  • bos taurus (domestic cattle)
KeywordsSIGNALING PROTEIN / Kinase / Regulator

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