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Comparison of Michaelis–Menten plots of HK853 autophosphorylation with Probes O, S, and N. HK853 (2 μM) was incubated with various concentrations of the probes (1 to 60 μM) at 25 °C. The reactions were quenched at 15 min, and TAMRA-N3 was conjugated by CuAAC to the propargyl handle (30 min). Formation of the pHis species was quantified by gel-based analysis (n = 2, error bars are standard error of the mean).
