PXD061315

PXD061315 is an original dataset announced via ProteomeXchange.

Title	Multiplicity of regulatory subunit conformations define structural ensemble of reset Protein Kinase A holoenzyme
Description	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 signal termination leading to 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 holoenzyme that capture an ensemble of conformational end-states through the integration of electron microscopy with structural mass spectrometry.Together, integrative cryo-EM with mass spectrometry showcases the high domain-specific dynamics of the regulatory subunit (RIα) and their interactions with Cα. Our integrated structure and dynamics approaches reveal that the tetrameric cAMP-free reset holoenzyme adopts multiple, distinct conformations of the RIα with contributions from the N-terminal linker and CNB-B domains. Our findings highlight the interplay between RIα, Cα and PDEs (PDE8) in signalosomes and the impact of substrate and nucleotides and offer a new paradigm for PDE-mediated regulation of cAMP-PKA signaling.
HostingRepository	PRIDE
AnnounceDate	2025-07-14
AnnouncementXML	Submission_2025-07-14_14:38:24.921.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Varun Venkatakrishnan
SpeciesList	scientific name: Bos taurus (Bovine); NCBI TaxID: 9913; scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue
Instrument	Orbitrap Eclipse; Q Exactive UHMR; Synapt MS

Revision	Datetime	Status	ChangeLog Entry
0	2025-02-28 10:01:29	ID requested
⏵ 1	2025-07-14 14:38:25	announced

Venkatakrishnan V, Laremore TN, Buckley TSC, Armache JP, Anand GS, Multiplicity of Regulatory Subunit Conformations Defines Structural Ensemble of Reset Protein Kinase A Holoenzyme. J Am Chem Soc, 147(17):14174-14190(2025) [pubmed]

10.1021/jacs.4c16269;

submitter keyword: Reset, Conformational dynamics, Signal termination, Phosphodiesterase,Protein Kinase A

Ganesh Srinivasan
contact affiliation	Department of Chemistry, Pennsylvania State University
contact email	gsa5089@psu.edu
lab head
Varun Venkatakrishnan
contact affiliation	Dept. of Chemistry, Pennsylvania State University
contact email	vxv5106@psu.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD061315
     1. Label: PRIDE project
     2. Name: Multiplicity of regulatory subunit conformations define structural ensemble of reset Protein Kinase A holoenzyme