TABLE 3.
Dissecting contribution of Ubp6 domains to kinetics of Rpn1 association
Full-length Ubp6 as well as isolated truncated UBL or ΔUBL domains were injected at various concentrations over immobilized full-length Rpn1. Values of kinetic parameters and of the KD dissociation constant were extracted from a fit of response curves to a minimal binding site model (single site for the individual domains or a two-site binding model for the full length protein).
| Protein | kon | koff | KD | |
|---|---|---|---|---|
| m−1s−1 | s−1 | μm | ||
| Ubp6 | Site 1 | 1.55 ± 0.36 × 104 | 9.61 ± 3.67 × 10−4 | 0.062 ± 0.004 |
| Site 2 | 3.53 ± 0.73 × 104 | 0.067 ± 0.006 | 1.92 ± 0.26 | |
| Ubp6ΔUBL | 1.81 ± 0.04 × 103 | 6.55 ± 0.95 × 10−4 | 0.36 ± 0.05 | |
| Ubp6UBL | 5.37 ± 0.54 × 104 | 0.120 ± 0.013 | 2.23 ± 0.24 | |