PXD035574

PXD035574 is an original dataset announced via ProteomeXchange.

Title	Structural insightsStructural insights into regulation of the PEAK3 pseudokinase scaffold by 14-3-3 into regulation of the PEAK3 pseudokinase scaffold by 14-3-3
Description	The three members of the PEAK family of pseudokinases (PEAK1, PEAK2, and PEAK3) are molecular scaffolds that have recently emerged as important nodes in signaling pathways that control cell migration, morphology, and proliferation, and are increasingly found mis-regulated in human cancers. While no structures of PEAK3 have been solved to date, crystal structures of the PEAK1 and PEAK2 pseudokinase domains revealed their dimeric organization. It remains unclear how dimerization plays a role in PEAK scaffolding functions as no structures of PEAK family members in complex with their binding partners have been solved. Here, we report the cryo-EM structure of the PEAK3 pseudokinase, also adopting a dimeric state, and in complex with an endogenous 14-3-3 heterodimer purified from mammalian cells. Our structure reveals an asymmetric binding mode between PEAK3 and 14-3-3 stabilized by one pseudokinase domain and the Split HElical Dimerization (SHED) domain of the PEAK3 dimer. The binding interface is comprised of a canonical primary interaction involving two phosphorylated 14-3-3 consensus binding sites located in the N-terminal domains of the PEAK3 monomers docked in the conserved amphipathic grooves of the 14-3-3 dimer, and a unique secondary interaction between 14-3-3 and PEAK3 that has not been observed in any previous structures of 14-3-3/client complexes. Disruption of these interactions results in the relocation of PEAK3 to the nucleus and changes its cellular interactome. Lastly, we identify Protein Kinase D as the regulator of PEAK3/14-3-3 binding, providing a mechanism by which the diverse functions of the PEAK3 scaffold might be fine-tuned in cells.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_09:02:45.061.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Antoine Forget
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue
Instrument	Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2022-07-25 02:30:50	ID requested
1	2023-07-20 10:49:52	announced
⏵ 2	2023-11-14 09:02:45	announced	2023-11-14: Updated project metadata.

Torosyan H, Paul MD, Forget A, Lo M, Diwanji D, Paw, ł, owski K, Krogan NJ, Jura N, Verba KA, Structural insights into regulation of the PEAK3 pseudokinase scaffold by 14-3-3. Nat Commun, 14(1):3543(2023) [pubmed]

submitter keyword: Human, AP-MS, LC-MSMS, PEAK3

Nevan Krogan
contact affiliation	UCSF
contact email	nevan.krogan@ucsf.edu
lab head
Antoine Forget
contact affiliation	University of California San Francisco
contact email	antoine.forget@ucsf.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD035574
     1. Label: PRIDE project
     2. Name: Structural insightsStructural insights into regulation of the PEAK3 pseudokinase scaffold by 14-3-3 into regulation of the PEAK3 pseudokinase scaffold by 14-3-3