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. 2015 Sep 10;10(11):2650–2657. doi: 10.1021/acschembio.5b00346

Figure 6.

Figure 6

The production of ATP by pyruvate kinase, monitored by Rho-MatB. (A) Time courses of fluorescence change upon ATP production by pyruvate kinase at different phosphoenolpyruvate concentrations: see Methods for details. The initial rates were determined by linear regression, using the slope obtained from a linear calibration (inset). (B) The parameters Km (75.5 ± 3.4 μM) and Vmax (0.019 ± 0.002 μM s–1) were obtained from a curve fit to the Michaelis–Menten model. The average Km and Vmax values (n = 5) are 103 ± 10 μM and 0.0190 ± 0.0004 μM s–1. This gives an average specific activity of 0.76 ± 0.02 μM s–1 U–1 mL. (C) Time courses of ATP production by pyruvate kinase as monitored by Rho-MatB in a stopped-flow apparatus: see Methods for details. Traces are offset by 0.20 μM ATP from each other for clarity. Rates were determined by linear regression and were plotted versus pyruvate kinase concentration (inset).