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

ABSTRACT: 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.

INSTRUMENT(S): Orbitrap Exploris 480

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

SUBMITTER: Evan Cornett  

PROVIDER: MSV000095132 | MassIVE | Mon Jun 24 11:58:00 BST 2024

SECONDARY ACCESSION(S): PXD053369

REPOSITORIES: MassIVE