Samples were reduced in loading buffer and separated by SDS-PAGE for 3-4 min followed by in-gel digestion with trypsin. Digests were analyzed by LC-MS/MS using a Waters nanoACQUITY interfaced to a Thermo Fusion Lumos MS. The LC separation 90 min gradient of 5-30% MeCN in a trapping configurations with a 75 um x 25 cm analytical column (Waters HSS T3). MS used orbitrap MS1 and iontrap MS/MS (OT-IT). Raw files were converted to .mgf using Proteome Discoverer and searched against a Swissprot human database appended with p53beta sequence and containing reverse decoys, with fixed carbamidomethyl modification on Cys and variable deamidation (NQ), oxidation (M), peptide N-terminal Gln-->pyroGlu and protein N-terminal acetylation. Data was annotated at a 1% peptide and protein FDR using Scaffold.
[doi:10.25345/C5678T]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: p53 ; BioID ; BirA ; GFP
Principal Investigators: (in alphabetical order) |
Michael Kastan, M.D., Ph.D., Duke University, USA |
Submitting User: | mwfoster |
Chen J, Zhang D, Qin X, Owzar K, McCann JJ, Kastan MB.
DNA-Damage-Induced Alternative Splicing of p53.
Cancers (Basel). Epub 2021 Jan 12.
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