<<< Full experiment listing

PXD034028-1

PXD034028 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleActivation loop phosphorylation tunes conformational dynamics underlying Pyk2 tyrosine kinase activation
DescriptionPyk2 is a multidomain non-receptor tyrosine kinase that undergoes a complex, multi-stage activation process. Ca2+-flux induces conformational rearrangements that relieve autoinhibitory FERM domain interactions. The kinase domain phosphorylates a key linker residue to recruit Src kinase. Pyk2 and Src mutually phosphorylate activation loop residues to confer full activation. While the mechanisms of autoinhibition are established, the conformational dynamics associated with autophosphorylation and Src recruitment remain unclear. Here, we employ hydrogen/deuterium exchange mass spectrometry (HDX-MS) to map the conformational changes associated with Src-mediated activation segment phosphorylation in a Pyk2 construct encompassing FERM and kinase domains (residues 20-692). Results reveal increased dynamics at regulatory interfaces spanning FERM and kinase domains. Phosphorylation of the activation segment stabilizes two antiparallel beta strands linking activation and catalytic loops. Increased dynamics of the C-terminal end of the activation loop propagate to the F-helix, explaining how phosphorylation prevents reversion to the autoinhibitory FERM interaction. HDX-guided site-directed mutagenesis and kinase activity profiling establish a mechanism for phosphorylation-induced active site sculpting to confer high activity.
HostingRepositoryPRIDE
AnnounceDate2023-03-08
AnnouncementXMLSubmission_2023-03-08_14:55:09.748.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterEricUnderbakke
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListphosphorylated residue
InstrumentSynapt MS
Dataset History
RevisionDatetimeStatusChangeLog Entry
02022-05-23 03:40:16ID requested
12023-03-08 14:55:10announced
22023-11-14 08:56:26announced2023-11-14: Updated project metadata.
Publication List
Keyword List
submitter keyword: kinase, phosphorylation, HDX-MS
Contact List
Eric StevenUnderbakke
contact affiliationRoy J. Carver Dept. of Biochemistry, Biophysics, and Molecular Biology, Iowa State University, Ames, IA USA
contact emailesu@iastate.edu
lab head
EricUnderbakke
contact affiliationIowa State University
contact emailesu@iastate.edu
dataset submitter
Full Dataset Link List
Dataset FTP location
NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/03/PXD034028
PRIDE project URI
Repository Record List
[ + ]