Table 1. Single-Turnover DNA Cleavage Rate Constants (kobs) using 1 μM SgrAI dimer.
32P-labeled Reporter DNA (1 nM) | Added Unlabeled Activator DNA | kobs a (min-1) a | Percent Cleaved a , b | Relative Acceleration c |
---|---|---|---|---|
18–1 | 0 nM PC d | 0.094±0.015 | - | - |
10 nM PC d | 0.18±0.06 | - | 2 | |
100 nM PC d | 0.30±0.03 | - | 3 | |
250 nM PC | 12.6±0.8 | 74±4 | 130 | |
500 nM PC | 27±7 | 90±10 | 300 | |
1000 nM PC d | 22±7 | - | 230 | |
1000 nM 22–1 e | 1.1±0.4 | 12 | ||
1000 nM 22-1-3’S | 1.47±0.12 | - | 16 | |
40–1 | 0 nM PC | 0.236±0.006 | 68±1 | - |
10 nM PC | 0.37±0.01 | 71±1 | 1.5 | |
100 nM PC | 4.4±0.4 | 69±2 | 20 | |
250 nM PC | 11.0±0.3 | 71.6±0.4 | 50 | |
500 nM PC | 18.0±0.3 | 75.8±0.2 | 80 | |
1 μM PC | 16.3±1.7 | 76.3±0.4 | 70 | |
2 μM PC | 23±2 | 73.0±0.8 | 98 | |
40–1 | 1 nM 40–1 | 0.28±0.02 | 63±2 | - |
1 μM 40–1 | 12±1 | 29±3 | 40 | |
40-2A | 0 nM PC | 0.013±0.004 | 17.7±0.5 | - |
10 nM PC | 0.018±0.004 | 70±7 | 1 | |
100 nM PC | 0.039±0.005 | 63±2 | 3 | |
250 nM PC | 0.82±0.09 | 28±2 | 60 | |
500 nM PC | 6.1±0.6 | 53±1 | 500 | |
1 μM PC | 9±3 | 53±7 | 700 | |
2 μM PC | 10±3 | 38±3 | 800 | |
40-2A | 1 nM 40-2A | 0.004±0.003 | 34±12 | - |
1 μM 40-2A | 0.25±0.06 | 78±5 | 20 | |
40-2B | 100 nM PC | 0.0221±0.0009 | 54±3 | 2 f |
250 nM PC | 1.23±0.07 | 20±4 | 100 f | |
500 nM PC | 2.2±0.4 | 29±2 | 170 f | |
1 μM PC | 8.1±1.4 | 33±2 | 600 f | |
2 μM PC | 11.8±1.4 | 45±2 | 900 f | |
40-2B-Top | 0 nM PC | 0.0129±0.0004 | 69.7±0.2 | - |
1 μM PC | 9.3±1.6 | 55.2±0.3 | 700 | |
40-2B- Bottom | 0 nM PC | 0.0023±0.0009 | 46±17 | - |
1 μM PC | 17±4 | 50±1 | 7000 | |
40-NC-TA | 0 nM PC | 0.024±0.004 | 11±1 | - |
1 μM PC | 0.052±0.006 | 5.2±0.6 | 2 |
aValues presented at the average of three independent trials ± the standard deviation.
bPercent Cleaved is the percentage of DNA cleaved by SgrAI in the assay by the given rate constant (kobs). In cases where less than 100% is cleaved by the given rate constant, the remainder is cleaved by a nonaccelerated rate constant.
cRelative Acceleration is the ratio of kobs to that in the absence of added unlabeled DNA.
dFrom Park, et al., 2010[8].
e From Park, et al., 2010[10].
fRelative to cleavage of 40-2B-Top with no PC DNA.