Tocriscreen mini library

For the screening, 60 nl of compounds from the Tocriscreen Mini library (Tocris Bioscience) were transferred to 1536-well plates. The mixtures of CFP-TNKS2-ARC4 (100 nM) and YFP-TBM (200 nM) or TNKS2 SAM(E897K)-CFP (150 nM) and TNKS2 SAM(Y920A)-YFP (300 nM) were prepared in assay buffer (10 mM Bis–Tris-Propane pH 7.0, 3%(w/v) PEG20,000, 0.01%(v/v) Triton-X100 and 0.5 mM TCEP), and rFRET signal was measured at two excitation wavelengths 410 nm and 430 nm. Protein mixtures were transferred to the plates using Mantis liquid dispenser (Formulatrix).

The compounds libraries used in this study included: the LOPAC library consisting of 1,280 pharmacologically active compounds (Sigma-Aldrich, St Louis, MO, USA); the Enzo Screen-Well kinase inhibitor library consisting of 80 kinase inhibitors; the Enzo Screen-Well Bioactive lipid library consisting of 190 bioactive lipids (Enzo Life Sciences, Farmingdale, NY, USA); the Tocriscreen Mini library consisting of 1,120 biologically active compounds (Tocris Bioscience, Bristol, UK); the Spectrum Collection consisting of 2,320 compounds (MicroSource, Gaylordsville, CT, USA); the Prestwick Chemical library consisting of 1,200 compounds; the Prestwick Phytochemicals Library consisting of 320 phytochemical compounds; the Prestwick Natural Compounds Library consisting of 240 natural compounds (Prestwick Chemical, Parc dinnovation, France). For all libraries the stock solution is 10 mM dissolved in DMSO. Before the screen experiment they were freshly pre-diluted 1:50 in PBS buffer.