Table II.
Peptide substrate steady-state kinetic parameters with three peptide substrates
| SOS2 Mutant | Peptide Substrate | Kcat | Km | Kcat/Km |
|---|---|---|---|---|
| s−1 | μm | m−1 s−1 | ||
| SOS2T168D | p1 | 1.89 | 211 ± 3.9 | 0.89 × 104 |
| p2 | 2.30 | 119 ± 2.8 | 1.93 × 104 | |
| p3 | 2.76 | 99 ± 1.1 | 2.79 × 104 | |
| SOS2T168DΔF | p1 | 2.01 | 201 ± 3.1 | 1.00 × 104 |
| p2 | 2.26 | 111 ± 1.8 | 2.04 × 104 | |
| p3 | 2.62 | 95 ± 1.5 | 2.76 × 104 | |
| SOS2T168DΔ308 | p1 | 1.90 | 198 ± 5.1 | 0.96 × 104 |
| p2 | 2.32 | 145 ± 4.1 | 1.60 × 104 | |
| p3 | 2.51 | 113 ± 4.8 | 2.22 × 104 |
Phosphorylation of the peptide substrate p1, p2, or p3 by SOS2T168D, SOS2T168DΔF, or SOS2T168DΔ308 was measured at optimal concentration of Mn2+ (2.5 mm). The kinetic parameters were determined by varying [p1], [p2], and [p3] while holding ATP at 10 μm. Kcat/Km is presented here as a measure of overall enzyme efficiency for each peptide substrate. Values are the means ± sd from three separate experiments.