Table 2.
Rates of processing duplex substrates by TDGWT and AcK analogs. aRates are reported as mean ± standard error (n). b25 nM U:G substrate, 1000 nM TDG. Asterisk (*) denotes p<0.01 versus slow phase of 3Q by ANOVA and post-hoc Tukey test for pairwise comparisons for U:G 25 nM substrate.
| Enzyme | Substrate | kmax (min−1)a |
kmax relative to T:G |
kSTO (min−1)a,b , yield (%) |
kTO (min−1)a |
kTO relative to T:G |
kmax/kTO |
|---|---|---|---|---|---|---|---|
| TDGWT | T:G | 0.30 ± 0.03 (4) | 1 | 0.007 ± 0.002 (7) | 1 | 43 | |
| U:G | 4.2 ± 0.2 (3) | 14 | 4.1 ± 0.6*, 91 (4) | 0.14 ± 0.05 (5) | 20 | 30 | |
| 5-FU:G | 1000 ± 290 (3) | 3300 | 2.5 ± 1.0 (3) | 360 | 400 | ||
| TDGK59Q | T:G | 0.42 ± 0.01 (7) | 1 | 0.003 ± 0.001 (7) | 1 | 140 | |
| U:G | 4.4 ± 0.4 (5) | 10 | 3.6 ± 0.10*, 96 (3) | 0.04 ± 0.01 (7) | 10 | 150 | |
| 5-FU:G | 940 ± 130 (3) | 2200 | 3.4 ± 0.8 (6) | 100 | 280 | ||
| TDGK83Q | T:G | 0.36 ± 0.02 (4) | 1 | 0.002 ± 0.001 (9) | 1 | 180 | |
| U:G | 4.4 ± 0.3 (3) | 12 | 1.9 ± 0.2*, 92 (3) | 0.09 ± 0.03 (9) | 45 | 50 | |
| 5-FU:G | 1260 ± 320 (3) | 3500 | 2.2 ± 0.5 (3) | 1100 | 570 | ||
| TDGK84Q | T:G | 0.46 ± 0.05 (4) | 1 | 0.009 ± 0.002 (6) | 1 | 51 | |
| U:G | 4.2 ± 0.3 (3) | 9.1 | 3.6 ± 0.5*, 93 (4) | 0.03 ± 0.01 (4) | 3.3 | 140 | |
| 5-FU:G | 980 ± 280 (3) | 2100 | 3.4 ± 0.7 (3) | 110 | 290 | ||
| TDGK87Q | T:G | 0.42 ± 0.05 (4) | 1 | 0.006 ± 0.003 (5) | 1 | 70 | |
| U:G | 3.9 ± 0.2 (3) | 9.3 | 2.1 ± 0.3*, 90 (4) | 0.11 ± 0.04 (4) | 18 | 35 | |
| 5-FU:G | 790 ± 14 (3) | 1900 | 2.3 ± 0.4 (3) | 380 | 340 | ||
| TDG3Q | T:G | 0.43 ± 0.05 (4) | 1 | 0.004 ± 0.002 (3) | 1 | 110 | |
| U:G | 3.1 ± 0.3 (5) | 7.2 | 2.9 ± 0.2*, 75 (3) | 0.05 ± 0.01 (7) | 13 | 62 | |
| 0.04 ± 0.01, 25 (3) | |||||||
| 5-FU:G | 750 ± 130 (3) | 1700 | 4.5 ± 0.9 (4) | 1100 | 170 |