Extended Data Table 3. Bio-layer interferometry analysis of SHOC2 ternary complex.
Binding kinetic values for step-specific complex member binding (Interaction) are indicated. Association rate constant (ka), dissociation rate constant (kd), steady-state dissociation constant (KD), and least squares fit (LSF) are shown. Values represent the mean of three replicates ± standard deviation. *Kinetics too irreversible to be determined.
| Interaction | SHOC2 | PP1C | MRAS | ka (1/Ms) x104 |
kd (1/s) x10−3 |
KD kinetic (M) x10−8 |
KD (M) x10−8 |
LSF (KD) x10−7 |
|---|---|---|---|---|---|---|---|---|
| SHOC2 >► PP1CA | WT | WT | — | 2.7 ± 0.1 | 390 ± 22 | 1,413 ± 135 | 670 | 5 |
| SHOC2 > PP1CAP50R | WT | P50R | — | 5.5 ± 1.4 | 182 ±79 | 96 ± 141 | 180 | 2.4 |
| SHOC2M1731 >■ PP1CA | M173I | WT | — | 5.0 ±0.7 | 384 ± 39 | 775 ± 137 | 450 | 3.2 |
| SHOC2M1731 >► PP1CAP50R | M173I | P50R | — | 7.0 ± 0.4 | 194 ± 46 | 279 ± 13 | 190 | 1.5 |
| SHOC2-PP1CA >■ MRAS | WT | WT | WT | 7.2 ± 0.9 | 5.2 ± 3.3 | 7.4 ± 0.7 | 20 | 0.6 |
| SHOC2-PP1CA >► MRASQ71L | WT | WT | Q71L | 8.1 ± 1.5 | 0.3 ± 0.1 | 0.4 ± 0.1 | 6.7 | 0.2 |
| SHOC2M173I-PP1CA >► MRAS | M173I | WT | WT | 6.4 ± 1.2 | 1.5 ± 1.3 | 2.4 ± 0.3 | 8.5 | 0 |
| SHOC2-PP1CAP50R >► MRAS | WT | P50R | WT | 7.9 ± 0.6 | 1.4 ± 0.8 | 1.8 ± 0.2 | 27 | 0.9 |
| SHOC2M173I-PP1CAP50R > MRAS | M173I | P50R | WT | 9.3 ± 1.4 | 0.5 ± 0.1 | 0.6 ± 0.1 | 20 | 1 |
| SHOC2M173I-PP1CAP50R > MRASQ71L | M173I | P50R | Q71L | * | * | * | 0.6 | 0.3 |
| SHOC2 >► PP1CB | WT | WT | — | 1.0 ± 4.4 | 7.5 ± 0.8 | 4,180 ± 2,570 | 1,100 | 6.2 |
| SHOC2-PP1CB >► MRAS | WT | WT | WT | 5.7 ± 6.9 | 2.8 ± 0.2 | 4.9 ± 0.5 | 20 | 0.6 |
| SHOC2 >► PP1CC | WT | WT | — | 3.5 ± 1.0 | 377 ± 41 | 1130 ± 112 | 650 | 12 |
| SHOC2-PP1CC >► MRAS | WT | WT | WT | 5.5 ± 5.6 | 3.7 ± 0.3 | 6.9 ± 0.6 | 19 | 0.4 |