Table 2. Kinetic Parameters for Inhibition of hBChE Variants by Nerve Agent Model Compoundsa.
kib (μM–1 min–1) | k2c (min–1) | KDd (μM) | t1/2 (min)e | R2f | |
---|---|---|---|---|---|
SPGB3N | |||||
wild type | 0.14 ± 0.08 | 1.17 ± 0.31 | 8.4 ± 3.9 | 3.8 ± 2.5g | 0.97 |
Y332S | 0.01 ± 0.01 | 0.52 ± 0.09 | 50.9 ± 11 | 9.5 ± 3.0g | 0.99 |
D340H | 0.03 ± 0.02 | 0.48 ± 0.04 | 19.0 ± 2.6 | 4.9 ± 1.3g | 0.99 |
Y332S/D340H | 0.002 ± 0.001 | 0.22 ± 0.35 | 132.0 ± 23.3 | 58 ± 2.16g | 0.94 |
G117H | 3.62 ± 3.03 | 0.02 ± 0.003 | 6202 ± 833 | NAh | 0.99 |
SPGD3N | |||||
wild type | 0.11 ± 0.05 | 0.58 ± 0.11 | 5.2 ± 2.2 | 4.0 ± 2.4g | 0.96 |
Y332S | 0.02 ± 0.01 | 0.24 ± 0.16 | 10.9 ± 1.18 | 22.6 ± 1.57g | 0.85 |
D340H | 0.04 ± 0.02 | 0.20 ± 0.08 | 5.4 ± 0.42 | 16.2 ± 1.07g | 0.84 |
Y332S/D340H | 0.03 ± 0.01 | 0.03 ± 0.004 | 1.1 ± 0.5 | 38.9 ± 7.5g | 0.76 |
G117H | 71.4 ± 5.59 | 0.19 ± 0.09 | 2613 ± 1500 | NAh | 0.97 |
SPGF3N | |||||
wild type | 2.03 ± 1.21 | 0.23 ± 0.01 | 0.11 ± 0.014 | 21.1 ± 8.97g | 0.99 |
Y332S | 0.71 ± 0.35 | 0.04 ± 0.01 | 0.06 ± 0.015 | 43.0 ± 16.2g | 0.99 |
D340H | 0.91 ± 0.45 | 0.05 ± 0.002 | 0.06 ± 0.005 | 48.6 ± 2.82g | 0.99 |
Y332S/D340H | 0.31 ± 0.04 | 0.27 ± 0.05 | 0.88 ± 0.124 | 42.4 ± 9.85g | 0.92 |
G117H | 4.3 ± 3.49 | 0.02 ± 0.02 | 3460 ± 7385 | NAh | 0.87 |
Values for k2 and KD were derived from the nonlinear curve fit of kapp over nerve agent model compound concentrations. Values reported represent best fit values ± the standard deviation derived from fitting 30–40 data points from a single experiment. Values for ki were calculated from k2/KD ratios. Statistical significance was evaluated by ANOVA followed by a Dunnett’s multiple-comparison test.
Bimolecular inhibition constant (ki).
Phosphonylation constant (k2).
Equilibrium dissociation constant (KD).
Half-life value for 50% inhibition of BTC hydrolysis.
Correlation constant for a nonlinear curve fit (R2).
Half-life values calculated for five concentrations of nerve agent model compounds.
Half-life value could not be calculated.