Description
We developed an ultra-deep O-GlcNAc proteomics workflow, by integrating multiple-proteases digestion, two mass spectrometric approaches (i.e., EThcD fragmentation and HCD product-dependent EThcD fragmentation), and two data analysis tools (i.e., MaxQuant and Proteome Discoverer). The performance of this strategy was benchmarked by the analysis of whole lysates from PANC-1 (a pancreatic cancer cell line), in total 2831 O-GlcNAc sites were unambiguously identified, representing the largest O-GlcNAc dataset of an individual study reported so far. Unexpectedly, in addition to confirming known sites and discovering many novel sites of Ser/Thr modification, O-GlcNAcylation was found on 121 tyrosine (Tyr) residues of 93 proteins. Through in vitro enzymatic assays, we revealed that OGT shows catalytic capacity to transfer O-GlcNAc onto Tyr residues of peptides and OGA can remove Tyr O-GlcNAcylation from peptides. Taken together, we discovered widespread O-GlcNAcylation on tyrosine residues of proteins and Tyr O-GlcNAcylation is mediated by OGT and OGA. As a novel form of glycosylation, Tyr O-GlcNAcylation may have important regulatory roles.
[doi:10.25345/C5ZP3WB4D]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: O-GlcNAc ; proteomics ; tyrosine ; OGT ; OGA
Contact
Principal Investigators:
(in alphabetical order)
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Junfeng Ma, Georgetown Univeristy, United States
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Submitting User: |
Chunyan
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Distinct protein accessions are counted across all files submitted in the "Statistical Analysis
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Distinct protein accessions are counted across all files submitted in the "Statistical Analysis
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"N/A" means no results of this type were submitted.
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