PXD064891

PXD064891 is an original dataset announced via ProteomeXchange.

Title	PERK protein kinase facilitates keratinocyte collective cell migration by engagement with cell adhesion molecules independent of its kinase activity
Description	Proximity labeling experiments (BioID) were performed on Human NTERT keratinocytes depleted of PERK (EIF2AK3, PERK-KO) re-expressing a WT PERK containing a C-terminal UltraID tag. PERK-KO and PERKUltraID cells were treated with differentiation medium containing 50 uM of exogenous Biotin for 6 h, washed three times with ice-cold PBS and harvested on ice with a modified RIPA buffer solution (25mM Tris-HCl, pH 7.5, 150 mM NaCl, 1% NP-40, 0.5% Sodium Deoxycholate, 1mM EDTA, 1x HALT protease/phosphatase inhibitor cocktail). Samples were then sonicated and incubated at 4C with rotation for 30 minutes, followed by centrifugation step to pellet cell debris. Total protein concentration was measured, and the samples were normalized to 1 mg/ml to a final volume of 1 ml. Normalized samples were incubated with 40uL of trypsin-resistant streptavidin beads with rotation overnight at 4C. The next day, the beads were collected by magnetic separation and washed three times with an IP wash solution (25 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.5% Triton X-100), followed by two washes with PBS. Finally, the beads were resuspended with 100 ul of PBS and submitted on ice for preparation and mass spectrometry analysis by the Indiana University School of Medicine Center for Proteome Analysis.
HostingRepository	MassIVE
AnnounceDate	2025-09-28
AnnouncementXML	Submission_2025-09-28_11:12:10.139.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Emma Doud
SpeciesList	scientific name: Homo sapiens; common name: human; NCBI TaxID: 9606;
ModificationList	Oxidation
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2025-06-11 12:14:00	ID requested
⏵ 1	2025-09-28 11:12:10	announced

Diaz MB, Staschke KA, Baucum AJ, Spandau DF, Wek RC, PERK protein kinase facilitates keratinocyte collective cell migration by engagement with cell adhesion molecules, independent of its kinase activity. Mol Biol Cell, 36(11):ar40(2025) [pubmed]

submitter keyword: BioID, MS, proteomics, NTERT, DatasetType:Proteomics

Dan F. Spandau
contact affiliation	Indiana University School of Medicine
contact email	dspanda@iu.edu
lab head
Amber L. Mosley
contact affiliation	Indiana University School of Medicine
contact email	almosley@iu.edu
lab head
Ronald C. Wek
contact affiliation	Indiana University School of Medicine
contact email	rwek@iu.edu
lab head
Emma Doud
contact affiliation	IUSM
contact email	edoud@iu.edu
dataset submitter

MassIVE dataset URI

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