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Protocol for mass spectrometric profiling of lysine malonylation by lysine acetyltransferase in CRISPRi K562 cell lines.
- Source :
-
STAR protocols [STAR Protoc] 2024 Jun 21; Vol. 5 (2), pp. 103074. Date of Electronic Publication: 2024 May 20. - Publication Year :
- 2024
-
Abstract
- Lysine malonylation is a protein posttranslational modification. We present a protocol to generate stable gene-knockdown K562 cell lines through lentiviral infection of a CRISPR interference (CRISPRi) system followed by lysine malonylation measurement using mass spectrometry (MS). We detail guide RNA (gRNA) vector cloning, lentiviral infection, cell line purification, protein digestion, malonyl-lysine enrichment, desalting, and MS acquisition and analysis. For complete details on the use and execution of this protocol, please refer to Zhang et al. <superscript>1</superscript> and Bons et al. <superscript>2</superscript> .<br />Competing Interests: Declaration of interests E.V. is a scientific co-founder of Napa Therapeutics and serves on the scientific advisory board of Seneque.<br /> (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
Details
- Language :
- English
- ISSN :
- 2666-1667
- Volume :
- 5
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- STAR protocols
- Publication Type :
- Academic Journal
- Accession number :
- 38771695
- Full Text :
- https://doi.org/10.1016/j.xpro.2024.103074