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. Author manuscript; available in PMC: 2009 Dec 4.
Published in final edited form as: Biochemistry. 2009 Feb 10;48(5):981–994. doi: 10.1021/bi801810h

Figure 2.

Figure 2

Dependence of the rate of formation of 3-PKB–ThDP and enamine on the concentration of 3-PKB. (A) Concentration dependence of the PKB–ThDP formation. BFDC (concentration of active centers 33.2 μM) in buffer B was mixed with an equal volume of 2, 4, 6, 8, and 10 mM 3-PKB in the same buffer at 30 °C. The reaction was monitored in the 300–595 nm wavelength range for 264 s. (B) Concentration dependence of enamine formation. Conditions as in A. Solid lines represent the best fit of 1/τ1 vs [3-PKB] and 1/τ2 vs [3-PKB] according to the equations 1/τ1 = k2[3-PKB]/(Kd1 + [3-PKB]) + k−2 and 1/τ2 = k3[3-PKB]/(Kd2 + [3-PKB]), respectively.