Table 1.
tRNA domain | Mutants of AatRNA | kcat (s−1) | Km (μM) | kcat/Km (μM−1s−1) | kcat/Km (relative) |
---|---|---|---|---|---|
native | 1.28 ± 0.34 | 1.10 ± 0.26 | 1.16 | 1.0 | |
Acceptor-arm | A73G | 0.06 ± 0.015 | 7.40 ± 2.6 | 0.008 | 0.007 |
A73U | 0.19 ± 0.05 | 7.65 ± 1.4 | 0.025 | 0.022 | |
A73C | 0.15 ± 0.06 | 6.29 ± 2.1 | 0.024 | 0.021 | |
A5:U68&G6:C67 | 1.46 ± 0.41 | 1.60 ± 0.45 | 0.91 | 0.78 | |
A4:U69&A5:U68 | 1.67 ± 0.33 | 0.83 ± 0.21 | 2.01 | 1.73 | |
C69U | 0.098 ± 0.014 | 3.29 ± 0.45 | 0.03 | 0.026 | |
D-arm | G13C | 0.56 ± 0.08 | 0.32 ± 0.056 | 1.75 | 1.51 |
A14U | 0.53 ± 0.12 | 18.00 ± 2.7 | 0.029 | 0.025 | |
A15U | 1.13 ± 0.15 | 1.21 ± 0.18 | 0.93 | 0.8 | |
C16G | 1.34 ± 0.31 | 1.05 ± 0.12 | 1.28 | 1.10 | |
U17A | 1.03 ± 0.26 | 1.18 ± 0.3 | 0.87 | 0.75 | |
G18C | 0.56 ± 0.062 | 5.20 ± 0.87 | 0.11 | 0.095 | |
G19C | nm | nm | nm | nm | |
C20A | 1.21 ± 0.17 | 1.50 ± 0.25 | 0.81 | 0.70 | |
A20aC | 0.94 ± 0.14 | 2.19 ± 0.38 | 0.43 | 0.37 | |
G21A | 0.59 ± 0.077 | 1.37 ± 0.22 | 0.43 | 0.37 | |
G13:C23 | 1.061 ± 0.32 | 3.76 ± 0.63 | 0.28 | 0.24 | |
G19U:C56A | 0.94 ± 0.13 | 1.49 ± 0.14 | 0.63 | 0.54 | |
Anticodon-arm | A35U | 1.52 ± 0.22 | 1.60 ± 0.21 | 0.95 | 0.82 |
A35C | 1.58 ± 0.37 | 0.94 ± 0.16 | 1.68 | 1.45 | |
A35G | 1.34 ± 0.24 | 0.97 ± 0.21 | 1.38 | 1.19 | |
Δ35-37 | 1.52 ± 0.32 | 0.77 ± 0.11 | 1.97 | 1.70 | |
ΔG29:C41 | 1.01 ± 0.21 | 0.89 ± 0.092 | 1.13 | 0.97 | |
C41U | 0.84 ± 0.12 | 0.85 ± 0.23 | 0.99 | 0.85 | |
ΔAC | 1.43 ± 0.19 | 2.77 ± 0.31 | 0.52 | 0.45 | |
Variable-arm | V-armSer | 0.11 ± 0.026 | 4.03 ± 0.55 | 0.027 | 0.023 |
V-arm-2bp | 0.75 ± 0.15 | 1.42 ± 0.26 | 0.53 | 0.46 | |
ΔV-arm | 0.38 ± 0.05 | 4.20 ± 0.57 | 0.09 | 0.078 | |
U48A | 0.64 ± 0.082 | 3.64 ± 0.53 | 0.18 | 0.16 | |
ΔG47k | 0.48 ± 0.05 | 4.02 ± 0.61 | 0.12 | 0.10 | |
ΔU48 | nm | nm | nm | nm | |
A47eU | 1.41 ± 0.15 | 0.85 ± 0.076 | 1.66 | 1.43 | |
TψC-arm | ΔC59 | nm | nm | nm | nm |
A58G | 1.57 ± 0.41 | 9.16 ± 1.86 | 0.17 | 0.15 |
All kinetic data were reproduced at least 3 times; nm: not measurable.