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Crosslinks in Soluble Extract of DSSO Crosslinked Intact Tetrahymena thermophila Cilia

ABSTRACT: Intact cilia were isolated from dibucaine-treated Tetrahymena thermophila and subjected to DSSO crosslinking. Following quenching of the DSSO, a soluble "membrane and matrix" extract was generated by the addition of buffer containing 1% NP-40. The insoluble axonemes were removed by centrifugation and the supernatant extract was digested with trypsin, and crosslinked peptides were enriched using size exclusion chromatography. Crosslinks were identified using an MS2-MS3 data collection method and the Proteome Discoverer XlinkX node. Crosslink searching used a smaller fasta generated from analysis of the .RAW files and the Basic Sequest workflow.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Tetrahymena Thermophila (ncbitaxon:5911)

SUBMITTER: Edward Marcotte

PROVIDER: MSV000090056 | MassIVE | Mon Aug 01 09:54:00 BST 2022

SECONDARY ACCESSION(S): PXD035702

REPOSITORIES: MassIVE

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