Table 2.
Enzymatic parameters are calculated by fitting the Michaelis-Menten equation to substrate titrations. Enzyme concentrations used for activity measurements were 400 nM, or if this caused saturated signals 50 nM (*).
DOI: http://dx.doi.org/10.7554/eLife.26640.012
Table 2—source data 1. Source data for values shown in Table 2.
All values are [PO4] (in μM). Numbers (#i) above data indicate independent experiment number. Most kinetic parameters (kcat, KM) are extracted from curves shown in Figure 3, for which source data are available with this figure. Below source data are shown for cases where experiments were repeated at higher enzyme concentration (400 nM) to extract reliable kinetic parameters. Equation used to extract Vmax and KM: Y = Bo + Vm*X/(X + KM); Variables: Vmax, KM, Bo = baseline. Software used: Graphpad Prism.
elife-26640-table2-data1.xlsx (52.8KB, xlsx)
DOI: 10.7554/eLife.26640.013
| Ptase | Ptase-C2 | |||||||
|---|---|---|---|---|---|---|---|---|
| IP4 | PI(3,4,5)P3 | IP4 | PI(3,4,5)P3 | |||||
| kcat (s−1) | KM (μM) | kcat (s−1) | KM (μM) | kcat (s−1) | KM (μM) | kcat (s−1) | KM (μM) | |
| WT | 1.32 ± 0.02 | 98 ± 7 | 0.69 ± 0.02 | 43 ± 4 | 2.02 ± 0.04 | 115 ± 8 | 7.83 ± 0.26* | 94 ± 9* |
| FLDD | 1.20 ± 0.05 | 82 ± 13 | 4.44 ± 0.17* | 61 ± 8* | 1.39 ± 0.06 | 73 ± 13 | 4.79 ± 0.49* | 122 ± 31* |
| R649A | 1.64 ± 0.05 | 126 ± 13 | 1.35 ± 0.06 | 59 ± 8 | 1.79 ± 0.05 | 110 ± 10 | 6.37 ± 0.42* | 100 ± 18* |