Table 1.
Rate constants for hydrolysis and aminolysis derived from nucleophile partitioning experiments
Kobs-HA (s−1) | Fold increase (HA) | Kobs-water (s−1) | Fold increase (water) | Fold RF defect | |
---|---|---|---|---|---|
Solution | 5.3×10−3 ± 0.7×10−3 | 19 | 2.3×10−5 | 2 | |
Ribosome | 2.9×10−4 ± 0.2×10−4 | 1.2×10−5 ± 0.4×10−5 | |||
tRNA | 1.3×10−3 ± 0.3×10−3 | 5 | 5.9×10−4 ± 2.3×10−4 | 50 | |
RF1 | cnd | 0.65 ± 0.06 | 53000 | ||
| |||||
Ribosome Mutants
| |||||
A2602C | 1.8×10−4 ± 0.4×10−4 | 1 | 1.0×10−3 ± 0.3×10−3 | 85 | 620 |
U2585C | 5.6×10−4 ± 0.3×10−4 | 2 | 8.2×10−3 ± 0.9×10−3 | 670 | 79 |
| |||||
RF1 Mutants
| |||||
G233A | 3.6×10−4 ± 2.4×10−4 | 1 | 1.2×10−3 ± 0.6×10−3 | 97 | 540 |
G234A | 2.9×10−4 ± 0.6×10−4 | 1 | 8.4×10−5 ± 2.6×10−5 | 7 | 7700 |
Q235W | cnd | 0.086 ± 0.003 | 7000 | 7 | |
Q235K | cnd | 0.17 ± 0.04 | 14000 | 4 | |
Q235E | cnd | 0.27 ± 0.05 | 22000 | 2 | |
Q235N | 2.1×10−5 ± 0.3×10−5 | −14 | 8.5×10−5 ± 2.0×10−5 | 7 | 7600 |
Q235D | 1.1×10−5 ± 0.1×10−5 | −26 | 6.8×10−5 ± 0.4×10−5 | 6 | 9500 |
Q235L | cnd | 0.0086 ± 0.0026 | 700 | 75 | |
Q235T | cnd | 0.016 ± 0.004 | 1300 | 41 | |
Q235S | 4.4×10−3 ± 0.6×10−3 | 15 | 0.068 ± 0.009 | 5500 | 10 |
Q235A | 3.3×10−2 ± 0.5×10−2 | 113 | 0.18 ± 0.02 | 14000 | 4 |
Q235G | 4.7×10−2 ± 0.8×10−2 | 165 | 0.021 ± 0.009 | 1700 | 30 |
cnd = could not determine