Table 1.
Steady state kinetic data for Kf-mediated insertion of modified triphosphates (dYTP) opposite their cognate nucleotide in the template (X).
| 5'–d(TAATACGACTCACTATAGGGAGA) | ||||
|---|---|---|---|---|
| 3'–d(ATTATGCTGAGTGATATCCCTCTXGCTAGGTTACGGCAGGATCGC) | ||||
|
| ||||
| X | Y | kcat (min−1) | KM (μM) |
kcat/KM × 105 (M−1
min−1) |
| T | Aa | 4.1 ± 0.3 | 0.0053 ± 0.0004 | 7700 |
| 5SICS | NaMa | 14.6 ± 0.8 | 0.25 ± 0.04 | 580 |
| MM02a | 14 ± 2 | 35.8 ± 0.6 | 4.0 | |
| MMO2PA | 11 ± 2 | 15 ± 2 | 7.2 | |
| MMO2A | 6.7 ± 0.3 | 69 ± 8 | 0.97 | |
| MMO2SSBIO | 1.5 ± 0.3 | 45 ± 10 | 0.34 | |
| NaM | 5SICSa | 8.3 ± 1.1 | 0.039 ± 0.004 | 2100 |
| 5SICSPA | 8.1 ± 0.9 | 1.0 ± 0.1 | 83 | |
| 5SICSA | 0.085 ± 0.05 | 19 ± 4 | 0.045 | |
| 5SICSαSc | 6.5 ± 1.5 | 0.42 ± 0.16 | 160 | |
| 5SICSBIO | 0.55 ± 0.13 | 31 ± 12 | 0.18 | |
| 5SICSSSBIO | 1.3 ± 0.3 | 29 ± 9 | 0.45 | |
a
Manuscript submitted.
b Below limit of detection.
c
Mixture of Sp and Rp diastereomers.