Table 1.
The mean time of different steps of elongation starting from either 70S IC or pre-T complex measured by quench-flow and stopped-flow
| A. Reaction starting from 70S IC, the elongation mix contains natural peptidyl tRNA | |||||||
|---|---|---|---|---|---|---|---|
| Quench-flow measurement |
Stopped-flow measurement | Derived mean times following the model shown in Figure 2A |
|||||
| mRNA | τp1 (ms) | τtripeptide (ms) | τp2 (ms) |
τfluor (ms) |
τtranslocation (τtripeptide – (τp1 + τp2)) (ms) | τmRNA mov (τfluor – τp1) (ms) |
τpost mRNA move (τtranslocation – τmRNA mov) (ms) |
| MFL+9 nodye | 32 ± 2 | 368 ± 20 | 100 ± 6 | - | 236 ± 15 | - | - |
| MFL+9 | 30 ± 2 | 315 ± 13 | 119 ± 15 | 79 ± 9 | 180 ± 18 | 49 ± 9 | 131 ± 9 |
| MFL+10 nodye | 32 ± 3 | 261 ± 15 | 75 ± 6 | - | 154 ± 16 | - | - |
| MFL+10 | 31 ± 3 | 244 ± 12 | 76 ± 10 | 83 ± 6 | 137 ± 16 | 52 ± 7 | 85 ± 16 |
| MFL+11 | 31 ± 3 | 255 ± 18 | 66 ± 8 | 93 ± 2 | 158 ± 20 | 62 ± 4 | 96 ± 20 |
| MFL+12 |
33 ± 3 |
217 ± 27 |
56 ± 6 |
- |
128 ± 28 |
- |
- |
| B. Reaction starting from pre-TC with NAc-Phe- tRNAPhe in the A sitef | |||||||
| |
Quench-flow measurement |
Stopped-flow measurement |
|||||
| mRNA |
τtranslocation + τp2 (ms) |
τmRNA mov (ms) |
|
|
|
|
|
| MFL+9 | 2381 ± 227 | 112 ± 6 | |||||
| MFL+10 | 1667 ± 139 | 119 ± 7 | |||||
See Materials and Methods and Figures 2A and 3A for derivation of the mean time parameters. The results are average of minimum three identical replicates with standard deviation.