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. 2019 Sep 11;9:13084. doi: 10.1038/s41598-019-49234-6

Figure 4.

Figure 4

Interaction and inhibition of PKM2 with SA and amoB1. (A,B) Thermal stability assay of PKM2 with different concentrations of SA (A) and amoB1 (B). For this assay, different concentrations of SA or amoB1 were incubated with 2 µM PKM2 for 30 min at RT, then melting curves were obtained by monitoring the fluorescence at 590 nm. (C) Trp intrinsic fluorescence assay of PKM2 with different concentrations of amoB1. For the Trp intrinsic fluorescence assay, different concentrations of amoB1 were incubated with 1 µM PKM2 for 30 min at RT, then Trp emission fluorescence was obtained (excitation/emission = 295 nm/350 nm). (D,E) Inhibition of PKM2 activity by SA (D) and amoB1 (E). For the inhibition assays, different concentrations of salicylates were incubated with 1.3 ng/uL PKM2 for 30 min at RT, then conversion of NADH to NAD+ was monitored (excitation/emission = 340 nm/460 nm). Data are mean ± SD (n = 3). (F) Effect of SA (0.4 mM) on PKM1 enzyme activity. Experiments were repeated three times with similar results.