Table 1. Kinetic Parameters for EcYihV in the Presence and Absence of Cellular Metabolites.
| variablea substrate | additive | kcat (s–1) | KM (mM) | KI (mM) | kcat/KM (s–1 mM–1) |
|---|---|---|---|---|---|
| ATPb,c | 3.1 ± 0.2 | 1.0 ± 0.2 | 3.2 ± 0.8 | ||
| SF | 1.2 ± 0.2 | 1.3 ± 0.3 | 8.5 ± 2.8 | 0.9 ± 0.4 | |
| SFd | 0.30 ± 0.5 | 0.3 ± 1.1 | |||
| SF | SQ | 0.7 ± 0.1 | 0.6 ± 0.2 | 16 ± 9 | 1.2 ± 0.4 |
| SF | SLA | 0.77 ± 0.13 | 0.60 ± 0.19 | 6.4 ± 2.7 | 1.3 ± 0.6 |
| SF | DHAP | 1.4 ± 0.3 | 0.61 ± 0.24 | 3.0 ± 1.4 | 2.3 ± 1.4 |
| SFc | citrate | 3.2 ± 0.4 | 1.6 ± 0.5 | 2.0 ± 0.9 | |
| SF | PEP | 1.9 ± 0.2 | 0.11 ± 0.04 | 4.4 ± 1.5 | 17.7 ± 8.6 |
| SF | F6P | 2.5 ± 1.2 | 0.97 ± 0.7 | 2.6 ± 1.9 | 2.5 ± 3.1 |
| SF | FBP | 1.2 ± 0.1 | 0.04 ± 0.01 | 5.8 ± 1.5 | 27.8 ± 10.8 |
| SFc | ADP | 0.14 ± 0.01 | 0.06 ± 0.03 | 2.3 ± 1.3 |
a
Reactions were conducted in 200 μL volume in 25 mM BTP buffer (pH 7.5), 25 mM KCl, 5 mM MgCl2, 0.1 mg/mL BSA, 36.6 nM EcYihV at 30 °C, for 60 min. [ATP] = 1.0 mM, and kinetic data were analyzed using a substrate inhibition kinetic model, unless otherwise noted. Additives were at 10 mM.
b
[SF] = 1.0 mM.
c
Kinetic parameters obtained by analysis using the Michaelis–Menten equation.
d
[ATP] = 0.1 mM.