Table 1.
ATP turnover numbers of human NLRP3 proteins.
| NLRP3 protein | turnover (min−1) | fold change relative to wt peak 1 |
|---|---|---|
| Native | ||
| wt (peak 1) | 0.385 | 1.00 |
| wt (peak 2) | 0.027 | 0.07 |
| Walker A | ||
| K232A | 0.132 | 0.34 |
| T233S | 0.464 | 1.21 |
| Walker B | ||
| D302A | 0.074 | 0.19 |
| G303E | 0.046 | 0.12 |
| D305A | 0.071 | 0.18 |
| E306Q | 0.086 | 0.22 |
| D305A/E306A | 0.084 | 0.22 |
| D305A/E306Q | 0.161 | 0.42 |
| G303E/D305A/E306A | 0.136 | 0.35 |
| Sensor 1 | ||
| R351T | 0.037 | 0.10 |
| R351G | 0.154 | 0.40 |
| Sensor 2 | ||
| H522D | 0.065 | 0.17 |
| H522R | 0.331 | 0.86 |
| M523A | 0.589 | 1.53 |
| P-motif | ||
| P412A | 0.077 | 0.20 |
| Glu-switch | ||
| R262W | 0.204 | 0.53 |
The ATP-hydrolysis reaction of full length MBP-NLRP3 and protein mutants (always peak 1) was analyzed over time by RP-HPLC using 3 µM NLRP3 protein, 100 µM ATP and 5 mM Mg2+ at 25 °C. For wild type NLRP3, peak 2 was also measured under the same conditions. The resulting turnover numbers (min−1) derived from the initial linear part of the curves are listed.