Probing the Role of O-GlcNAcylation in Modulating Protein-DNA Interactions: Insights from a Proteomic Transcription Factor Response Element-Based Pulldown Screening Approach

O-linked β-N-acetylglucosamine (O-GlcNAc), a critical post-translational modification (PTM) predominantly found in the nucleus, plays a substantial role in regulating gene expression by modulating transcription factors (TFs) activity. Herein, by utilizing a pulldown screening approach termed as tandem array of transcription factors response elements (catTFREs), we unravel the profound impact of fluctuating levels of O-GlcNAcylation on the DNA binding efficiency of endogenous TFs. The proteins were enriched for label-free quantitative proteomics by LC-MS/MS.