UNC6535 TFA is a covalent ligand targeting the triple Tudor domain (3TD) of SETDB1. UNC6535 TFA reversibly binds to the aromatic cages of both TD2 and TD3 within SETDB1 3TD simultaneously, weakly inhibits the methyltransferase activity of SETDB1, and displaces the H3K9Me2K14Ac peptide from SETDB1 3TD [1].
In Vitro, UNC6535 TFA reversibly binds to the purified tripartite Tudor domain of SETDB1 and displaces the H3K9Me2K14Ac peptide, with an IC50 of 3.4 μM[1]. UNC6535 TFA binds to the purified triple Tudor domain of SETDB1, with a measured Kd of 4.2 μM determined by isothermal titration calorimetry[1]. UNC6535 TFA inhibits the methyltransferase activity of purified full-length SETDB1 protein, with an IC50 of 84 μM[1]. UNC6535 TFA inhibits the methyltransferase activity of purified truncated SETDB1-S (lacking the triple Tudor domain) with an IC50 of 30 μM[1]. UNC6535 TFA displaces the H3K9Me2K14Ac peptide from purified truncated SETDB1-S (lacking the tripartite Tudor domain) via fluorescence polarization, with an IC50 of 8.6 μM[1].
References:
[1]. Uguen M, et al. Potent and selective SETDB1 covalent negative allosteric modulator reduces methyltransferase activity in cells. Nat Commun. 2025;16(1):1905. Published 2025 Feb 24.
In Vitro, UNC6535 TFA reversibly binds to the purified tripartite Tudor domain of SETDB1 and displaces the H3K9Me2K14Ac peptide, with an IC50 of 3.4 μM[1]. UNC6535 TFA binds to the purified triple Tudor domain of SETDB1, with a measured Kd of 4.2 μM determined by isothermal titration calorimetry[1]. UNC6535 TFA inhibits the methyltransferase activity of purified full-length SETDB1 protein, with an IC50 of 84 μM[1]. UNC6535 TFA inhibits the methyltransferase activity of purified truncated SETDB1-S (lacking the triple Tudor domain) with an IC50 of 30 μM[1]. UNC6535 TFA displaces the H3K9Me2K14Ac peptide from purified truncated SETDB1-S (lacking the tripartite Tudor domain) via fluorescence polarization, with an IC50 of 8.6 μM[1].
References:
[1]. Uguen M, et al. Potent and selective SETDB1 covalent negative allosteric modulator reduces methyltransferase activity in cells. Nat Commun. 2025;16(1):1905. Published 2025 Feb 24.