Dissociation constants of F64W-H122G and F64W mutant NDPKs for NDPs, NTPs, and nonhydrolyzable ATP analogs
| Nucleotide | F64W-H122G | F64W |
|---|---|---|
| KD (μM) | ||
| dTTP | 1.2 ± 0.1 | n.m. |
| ddTTP | 4.0 ± 0.6 | n.m. |
| AZT-TP | 30 ± 3 | n.m. |
| D4T-TP | 1.2 ± 0.1 | n.m. |
| ATP | 0.20 ± 0.05 | n.m. |
| GTP | 0.15 ± 0.05 | n.m. |
| CTP | 7.0 ± 1.0 | n.m. |
| AMP-PNP | 30 ± 3 | 60 ± 6 |
| AMP-PCP | 750 ± 100 | 1000 ± 200 |
| dTDP | 75 ± 20 | 100 ± 503-a |
| AZT-DP | 125 ± 10 | 80 ± 203-a |
| D4T-DP | 85 ± 10 | no signal |
| ADP | 25 ± 1 | 25 ± 53-a |
| GDP | 15 ± 2 | 14 ± 53-a |
| CDP | 210 ± 20 | 200 ± 1003-a |
Titrations of F64W-H122G NDPK with natural and anti-HIV nucleotides were performed as described in the legend to Figure 2. Data presented are the mean values of three determinations ± S.E.
n.m., nonmeasurable.
↵3-a The binding of NDPs to F64W is measured as shown inSchneider et al. (1998b).