Biopolym. Cell. 2025; 41(1):42-51.
http://dx.doi.org/10.7124/bc.000B0B
Molecular and Cell Biotechnologies
The protein kinase CK2 inhibitor testing by capillary electrophoresis
1Mahdysiuk M. V., 1, 2Bieda O. A., 1Bdzhola V. G., 1Volynets G. P., 3Karbovskyi L. L., 1Balanda A. O., 1Lukashov S. S., 1Yarmoluk S. M.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03143
  2. LLC “Scientific and service firm “Otava”
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03143
  3. Biotech API Laboratory, JSC Farmak
    63, Frunze Str., Kyiv, Ukraine, 04080

Abstract

Aim. This research aimed to enhance the methodology for efficiently evaluating CK2 inhibitors using capillary electrophoresis and validate the techniques with the CK2 inhibitor FNH79. Additionally, the study included the assessment of a novel potential inhibitor, the aurone derivative BFO21, using the optimized protocol. Methods. The research was conducted with the capillary electrophoresis method, and 150 mM orthophosphoric acid (pH 1.2) as a background electrolyte. Conversion of enzymatic reaction was calculated as the ratio of the phosphorylated product peak area to the total peak area of both substrate and product. Results. The optimal testing conditions were determined, involving 50 units of the enzyme per 50 μl of the reaction mixture, an initial peptide substrate concentration of 100 μM, and an incubation time of 40 minutes. The initial concentration of ATP was 100 μM. FNH79 demonstrated IC50 and Ki values of 94 nM and 4.5 nM, respectively. The new aurone compound BFO21 exhibited IC50 and Ki values of 44 nM and 2.1 nM, respectively. Conclusions. Under optimized testing conditions, the activity values for the FNH79 inhibitor strongly matched previously published results. Additionally, the activity values of the BFO21 inhibitor revealed its significant potential as the CK2 inhibitor.
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