MassIVE MSV000095132

Complete Public PXD053369

Quantitative analysis of non histone lysine methylation sites and lysine demethylases in breast cancer cell lines

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Description

Tandem Mass Tag labeling (TMT labels) based mass spectrometry was used to quantify lysine methylation from four different cell lines (n=4 replicates): MCF7 (ER positive breast cancer), MDAMB231 (TNBC), HCC1806 (TNBC), and MCF10A (non-cancerous), sans antibody enrichment. To boost the detection of lysine demethylases, we generated a GFP-KDM Trigger Channel. Briefly, 27 GFP-KDMs were overexpressed and immunoprecipitated from HEK293T cells. The immunoprecipitated proteins were subsequently trypsin digested, TMT labeled, and added to the Breast Cancer 16-plex. All runs were conducted on the Orbitrap Exploris and processed using Proteome Discoverer. Database searching was conducted using Sequest HTS using the Uniprot-reviewed human proteome. Downstream data and bioinformatic analysis was conducted using R. [doi:10.25345/C54X54T68] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: lysine methylation ; Isobaric Trigger Channel ; lysine demethylases ; breast cancer

Contact

Principal Investigators:
(in alphabetical order) 		Evan Cornett, Indiana University School of Medicine, United States
Submitting User: causherm

Publications

Berryhill CA, Evans TN, Doud EH, Smith-Kinnaman WR, Hanquier JN, Mosley AL, Cornett EM.
Quantitative analysis of non-histone lysine methylation sites and lysine demethylases in breast cancer cell lines.
bioRxiv. Epub 2024 Sep 22.

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