Table 3.
Recombinant SARS-CoV Main Protease | Kinetic parameters |
Dissociation parameters |
||||
---|---|---|---|---|---|---|
Kma (μM) | kcatb (10−3 s−1) | kcat/Kmc (nM−1 s−1) | Kdd (μM) | koffe (s−1) | ΔGo′f(kcal mol−1) | |
WT | 11 ± 3 | 32 ± 3 | 3.0 ± 1 | 0.08 ± 0.005 | 0.01 ± 0.001 | −1.5 ± 0.09 |
Δ(306) | 21 ± 7 | 43 ± 6 | 2.1 ± 0.7 | 0.1 ± 0.004 | 0.6 ± 0.03 | −1.3 ± 0.05 |
Δ(305−306) | 14 ± 3 | 39 ± 0.4 | 2.7 ± 0.5 | 0.08 ± 0.005 | 0.14 ± 0.01 | −1.5 ± 0.09 |
Δ(304–306) | 6 ± 1 | 27 ± 1 | 4.6 ± 0.9 | 0.007 ± 0.001 | 0.01 ± 0.001 | −2.9 ± 0.2 |
Δ(303–306) | 4 ± 2 | 22 ± 3 | 5.9 ± 3 | 0.34 ± 0.02 | 0.08 ± 0.004 | −0.6 ± 0.04 |
Δ(302–306) | 22 ± 7 | 30 ± 4 | 1.4 ± 0.6 | 0.6 ± 0.02 | 0.01 ± 0.001 | −0.3 ± 0.01 |
Δ(301–306) | 30 ± 14 | 25 ± 6 | 0.8 ± 0.4 | 0.3 ± 0.01 | 0.1 ± 0.003 | −0.7 ± 0.02 |
Δ(300–306) | 15 ± 5 | 17 ± 3 | 1.1 ± 0.4 | 0.07 ± 0.003 | 0.05 ± 0.002 | −1.6 ± 0.07 |
Δ(299–306) | 13 ± 3 | 1 ± 0.1 | 0.08 ± 0.02 | 49 ± 4 | 2.3 ± 0.1 | 2.3 ± 0.2 |
Δ(298–306) | 30 ± 12 | 0.6 ± 0.1 | 0.02 ± 0.01 | 313 ± 22 | 36 ± 2.2 | 3.4 ± 0.2 |
Δ(297–306) | 21 ± 5 | 0.04 ± 0.004 | 0.002 ± 0.001 | 393 ± 39 | 123±9.8 | 3.5 ± 0.3 |
R298A | 11 ± 4 | 3.4 ± 0.7 | 0.3 ± 0.1 | 193 ± 10 | 63 ± 3 | 3.1 ± 0.2 |
R298K | 16 ± 6 | 17 ± 3 | 1.1 ± 0.5 | 1.5 ± 0.01 | 4.6 ± 0.02 | 0.2 ± 0.001 |
R298L | 13 ± 4 | 4.8 ± 0.7 | 0.4 ± 0.1 | 45 ± 6 | 1.1 ± 0.1 | 2.2 ± 0.3 |
R298A/Q299A | 10 ± 5 | 0.21 ± 0.1 | 0.02 ± 0.01 | 218 ± 9 | 61 ± 2.4 | 3.1 ± 0.1 |
Q299A | 18 ± 7 | 0.7 ± 0.1 | 0.04 ± 0.02 | 61 ± 9 | 8.7 ± 0.9 | 2.4 ± 0.4 |
Q299E | 5 ± 2 | 2.2 ± 0.2 | 0.4 ± 0.1 | 90 ± 14 | 0.2 ± 0.02 | 2.6 ± 0.4 |
Q299K | 37 ± 24 | 2.1 ± 0.7 | 0.06 ± 0.04 | 19 ± 1 | 5.1 ± 0.2 | 1.7 ± 0.1 |
Q299N | 5 ± 1 | 1.7 ± 0.1 | 0.4 ± 0.01 | 52 ± 6 | 1.2 ± 0.1 | 2.2 ± 0.1 |
S123A | 13 ± 7 | 33 ± 7 | 2.6 ± 1.4 | 0.8 ± 0.04 | 0.02 ± 0.001 | −0.1 ± 0.01 |
S123A/R298A | 12 ± 9 | 6.9 ± 2 | 0.6 ± 0.5 | 24 ± 2 | 3.0 ± 0.2 | 1.9 ± 0.2 |
S123C | 13 ± 5 | 35 ± 3 | 2.7 ± 1.0 | 0.9 ± 0.1 | 0.1 ± 0.01 | −0.06 ± 0.03 |
S139A | 10 ± 4 | 33 ± 3 | 3.2 ± 1.2 | 0.4 ± 0.01 | 0.2 ± 0.002 | −0.5 ± 0.01 |
S139A/Q299A | 21 ± 9 | 4.8 ± 1 | 0.2 ± 0.1 | 22 ± 1 | 1.1 ± 0.04 | 1.8 ± 0.1 |
Michaelis constant for the 11-mer peptide ABZ-TSAVLQSGFRK-DNP.
Catalytic constant for the 11-mer peptide ABZ-TSAVLQSGFRK-DNP.
Specificity constant for the 11-mer peptide ABZ-TSAVLQSGFRK-DNP.
Dissociation equilibrium constant of the system.
Dissociation rate constant of the system.
Dissociation Gibb’s free energy of the system.