Table 1.
ATPa (μM) | Time (min.)b (A&B) | Rate (μ | M/min)c | Time (min.) (A&B) | Rate (μM/min)d | |
---|---|---|---|---|---|---|
1A | 1B | 2A | 2B | |||
0 | 15.2e | 0 | 0 | 16.1 | 0 | 0 |
5 | NAf | NA | NA | 26.1 | −0.005 | −0.005 |
10 | NA | NA | NA | 37.4 | 0.125 | 0.030 |
10 | NA | NA | NA | 46.9 | 0.058 | 0.069 |
10 | NA | NA | NA | 56.9 | 0.080 | 0.089 |
15 | NA | NA | NA | 67.3 | 0.150 | 0.153 |
30 | NA | NA | NA | 78.1 | 0.237 | 0.158 |
65 | 25.1 | 0.322 | 0.343 | 90.3 | 0.341 | 0.354 |
140 | 36.4 | 0.351 | 0.345 | 100.2 | 0.362 | 0.362 |
287.5 | 46.0 | 0.443 | 0.413 | 111.1 | 0.453 | 0.434 |
567.5 | 56.0 | 0.419 | 0.524 | 122.7 | 0.468 | 0.593 |
1125 | 66.4 | 0.645 | 0.610 | 132.0 | 0.633 | 0.631 |
Each titration was performed in duplicate (A and B). The reactions contained a fivefold excess of the +8 loop DNA substrate over Msh2-Msh3.
Total incubation time when rate was calculated.
Calculated rates for titration 1, starting at 65μM ATP.
Calculated rates for titration 2, starting at 5 μM ATP.
The time for replicate A is shown. The time for replicate B is less than 30 seconds later. It takes the reaction approximately 15 minutes to equilibrate.
Not applicable – measurements were not taken at these ATP concentrations.