OSMI-4

Preparation Method

OGT reactions were carried out in a 50-µL final volume, containing 0.1 mM UDP-GlcNAc, 200 nM purified full-length OGT, OSMI-4a/b,100 µM RBL-2 peptide in OGT reaction buffer. Reactions were incubated at 37℃ for 2 h. Afterwards, each reaction was transferred in duplicate into a 96-well white microplate and was mixed with a 1:1 ratio of the UDP-Glo Detection Reagent. After incubation at room temperature for 1 h, the luminescence was recorded with a POLARstar® Omega microplate reader or with a BioTek Synergy™ H4 microplate reader.

Reaction Conditions

37℃ for 2 h

Applications

IC₅₀ of OSMI-4a and OMSI-4b are 1.5 and 0.5μM, respectively.

Cell lines

HEK293T

Preparation Method

Cells were plated and grown in their respective media until they reached a confluency of 60-80%. For HEK293T cells, media was changed 3 hours before OSMI-4 was added; OSMI-4 was added directly to each well at the indicated concentrations. The number of dead cells were assessed by CellToxTM Green cytotoxicity Assay.

Reaction Conditions

0.125,0.25,0.5,1,2,4,8,16μM，24 h

Applications

EC50 of OSMI-4 in cells is 3 μM

References:

[1]. Loi EM, Weiss M, Pajk S, Gobec M, Tomašič T, Pieters RJ, Anderluh M. Intracellular Hydrolysis of Small-Molecule O-Linked N-Acetylglucosamine Transferase Inhibitors Differs among Cells and Is Not Required for Its Inhibition. Molecules. 2020 Jul 25;25(15):3381.

[2]. Martin SES, Tan ZW, Itkonen HM, Duveau DY, Paulo JA, Janetzko J, Boutz PL, Törk L, Moss FA, Thomas CJ, Gygi SP, Lazarus MB, Walker S. Structure-Based Evolution of Low Nanomolar O-GlcNAc Transferase Inhibitors. J Am Chem Soc. 2018 Oct 24;140(42):13542-13545.