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Mass spectrometric profiling of lysine malonylation in CRISPRi 562 cells lines to investigate malonylation activity of lysine acetyltransferases

ABSTRACT: Lysine malonylation is a protein posttranslational modification with potentially broad biological impact. We present a protocol to generate stable gene knockdown K562 cell lines through lentiviral infection of a CRISPRi system followed by lysine malonylation measurement using mass spectrometry (MS). We detail gRNA vector cloning, lentiviral infection, cell line purification, protein digestion, malonyllysine enrichment, desalting, and MS acquisition and analysis. This protocol can be applied for CRISPRa cell line generation and lysine malonylation profiling in tissues, with slight changes.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Birgit Schilling

PROVIDER: MSV000093564 | MassIVE | Mon Dec 04 15:09:00 GMT 2023

SECONDARY ACCESSION(S): PXD047514

REPOSITORIES: MassIVE

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