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Analysis of Protein Cysteine Acylation Using a Modified Suspension Trap (Acyl-Trap)

ABSTRACT: Mouse brain and MLE-12 lysates were alkylated with N-ethylmaleimide, precipiated with methanol and trapped on a pyridyl disulfide quartx (PDQ) suspension trap. Protein were washed and incubated with -/+hydroxylamine and trypsin, and unbound peptides were labeled with TMTPro reagents on C18 StageTips. Bound peptides were labeled with TMTPro reagents on PDQ traps, eluted with DTT and alkylated with iodoacetamide followed by StageTip cleanup. Peptides were analyzed by nanoLC-MS/MS using a 90 min gradient in duplicate. MS/MS used a SPS-MS3 (or FAIMS-SPS-MS3) method as indicated in metadata. Raw files were converted to .mzml and analyzed using FragPipe.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Mus Musculus (ncbitaxon:10090)

SUBMITTER: Michael T. Forrester Matthew W. Foster

PROVIDER: MSV000094234 | MassIVE | Mon Mar 04 17:53:00 GMT 2024

SECONDARY ACCESSION(S): PXD050341

REPOSITORIES: MassIVE

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