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Journal: Nucleic Acids Research
Article Title: Molecular determinants for recognition of serotonylated chromatin
doi: 10.1093/nar/gkaf612
Figure Lengend Snippet: H3Q5ser stabilizes Taf3-PHD/H3K4me2 and Taf3-PHD/H3K4me3 complexes. ( A ) Experimental (blue) and fitted (red) spectra of free, ∼50% bound, and fully bound TAF3-PHD resonances E862 and C896 for H3K4me3/H3K4me3Q5ser complex. Transfer peaks indicated with *. Experimental spectrum of E862 is also shown separately for clarity. ( B ) ITC analysis of GST-tagged Taf3-PHD binding to H3K4me3 in yellow and H3K4me3Q5ser in navy blue. Top panel displays the raw heat change per injection of H3 peptide. Bottom panel represents the binding isotherm. The solid line represents the best fit to the one-set-of-sites model using MicroCal-ITC Origin 7 software.
Article Snippet: ITC data were subsequently analysed and fitted with the one-site binding model in the
Techniques: Binding Assay, Injection, Software
Journal: Nucleic Acids Research
Article Title: Molecular determinants for recognition of serotonylated chromatin
doi: 10.1093/nar/gkaf612
Figure Lengend Snippet: Taf3-PHD P881 and W894 form key interaction surface with the serotonin group. ( A ) ITC analysis of GST-tagged Taf3-PHD mutants, BPTF-PHD , PHF2-PHD, or PHF8-PHD with the peptides H3K4me3 (in yellow) and H3K4me3Q5ser (in blue). Top panels display the raw heat change per injection of H3 peptide. Bottom panels represent the binding isotherm. The solid line represents the best fit to the one-set-of-sites model using MicroCal-ITC Origin 7 software. ( B ) Multiple sequence alignment (MSA) of human PHD fingers for which high-resolution structures in complex with H3K4me3 are available (ASH1L PDB: 8VLF; BPTF(2) PDB: 2FUU; DIDO1 PDB: 4L7X; ING1 PDB: 2QIC; ING4 PDB: 2PNX/2VNF; ING5 PDB: 3C6W; KMT2E PDB: 4L58; PHF2 PDB: 3KQI; PHF13 PDB: 3O7A; UHRF1 PDB: 3SOW). Consensus Cys4–His–Cys3 positions are highlighted in grey and indicated on top. Locations of the secondary structure elements of TAF3-PHD are indicated on top. S880 and A902 are highlighted in purple, while P881 and W894 are highlighted in yellow.
Article Snippet: ITC data were subsequently analysed and fitted with the one-site binding model in the
Techniques: Injection, Binding Assay, Software, Sequencing