MassIVE MSV000093453

Partial Public

O-GlcNAc Containing Proteins in Normal and Idiopathic Pulmonary Fibrosis Human Fibroblasts

Description

Isolated normal and IPF fibroblasts were homogenized in cold MilliQ water using a bullet blender. Samples were centrifuged and inhibitors were added: HALT (Thermo Fisher Scientific), Z-Pugnac (Tocris), Thiamet G (Cayman Chemicals), and benzonase (E1014, Millipore, Sigma). O-GlcNAc enzymatic labeling and protein capturing was performed as described using a Click-IT enrichment kit following the manufacturer's protocol (cat no: C33368, C33372, and C10416; Thermo Fisher Scientific). Following an alkyne agarose bead enrichment, proteins were reduced (5 mM DTT, 30 min at 37 C), alkylated (40 mM iodoacetamide, 1 h at 37C) and digested with trypsin (1:50 enzyme/protein ratio). 0.5 ug of samples were analyzed by LC-MS/MS on a Q-Exactive HF-X mass spectrometer. Data was searched using MSFragger in match between runs mode. [doi:10.25345/C5Z02ZK7J] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: fibroblasts ; O-GlcNAc ; fibrosis ; glycan

Contact

Principal Investigators:
(in alphabetical order)
Jennifer E. Kyle, Pacific Northwest National Laboratory, United States
Submitting User: alchemistmatt
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Originally identified proteins that were automatically remapped by MassIVE to proteins in the SwissProt human reference database.

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Distinct protein accessions are counted across all files submitted in the "Statistical Analysis of Quantified Analytes" category having a "Protein" column in this dataset.

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