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PXD033063

PXD033063 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleSpatiotemporal proximity labeling tools to track GlcNAc-modified functional protein hubs during cellular signaling
DescriptionA fundamental mechanism that all eukaryotic cells use to adapt to their environment is dynamic protein modification with monosaccharide sugars. In humans, O-linked N-acetylglucosamine (O-GlcNAc) is rapidly added and removed on diverse protein sites as a response to fluctuating nutrient levels, stressors, and signaling cues. Two aspects elude methods to track O-GlcNAc response functions with chemical strategies: spatial control over subcellular locations and time control during labeling reactions. The objective of this study was to create intracellular proximity labeling tools to identify functional changes of O-GlcNAc patterns with spatiotemporal control. We developed a labeling strategy based on TurboID system for rapid protein biotin conjugation that was directed to O-GlcNAc protein modifications inside cells, a set of tools we call “GlycoID.” Localized variants to the nucleus and cytosol, nuc-GlycoID and cyt-GlycoID, label O-GlcNAc proteins and their interactomes in subcellular space. Labeling during insulin as well as serum stimulation revealed functional changes in O-GlcNAc proteins in as soon as 30 minutes of signaling. We demonstrate that the GlycoID strategy can capture O-GlcNAcylated “activity hubs” consisting of O-GlcNAc proteins and their associated protein-protein interactions. The ability to follow changes in O-GlcNAc hubs during physiological events like insulin stimulation poises these tools to be used for determining mechanisms of glycobiological cell regulation. Our datasets in HeLa cells will be a useful functional resource for O-GlcNAc-associated physiology.
HostingRepositoryPRIDE
AnnounceDate2023-11-14
AnnouncementXMLSubmission_2023-11-14_08:27:29.648.xml
DigitalObjectIdentifierhttps://dx.doi.org/10.6019/PXD033063
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportSupported dataset by repository
PrimarySubmitterCharlie Fehl
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListbiotinylated residue
InstrumentOrbitrap Fusion
Dataset History
RevisionDatetimeStatusChangeLog Entry
02022-04-07 03:29:41ID requested
12022-10-14 20:04:13announced
22023-11-14 08:27:30announced2023-11-14: Updated project metadata.
Publication List
10.6019/PXD033063;
Liu Y, Nelson ZM, Reda A, Fehl C, Spatiotemporal Proximity Labeling Tools to Track GlcNAc Sugar-Modified Functional Protein Hubs during Cellular Signaling. ACS Chem Biol, 17(8):2153-2164(2022) [pubmed]
Keyword List
submitter keyword: glycosylation, insulin signaling, proximity labeling, nucleocytoplasmic,O-GlcNAc
Contact List
Charlie Fehl
contact affiliationChemistry department, Wayne State University US
contact emailcharlie.fehl@wayne.edu
lab head
Charlie Fehl
contact affiliationWayne State University Chemistry
contact emailcharlie.fehl@wayne.edu
dataset submitter
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Dataset FTP location
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