Table 3.
SPR measurements of the interaction of immobilized GST-Hpr1 with His-UBA-Mex67 as an analyte in the absence or in the presence of Lys48-linked tetraubiquitin (K48-Ub4)
| Analyte | Apparent KD (M) | Rmax (RU) | k2 (s−1) | k−2 (s−1) |
|---|---|---|---|---|
| UBA (5 μM)+ | ||||
| 0 | 3.7 × 10−6 | 31 | 6.5 × 10−3 | 9.9 × 10−4 |
| K48-Ub4 (1 μM) | 4.3 × 10−6 | 83 | 6.7 × 10−3 | 9.8 × 10−4 |
| K48-Ub4 (3.5 μM) | 5.0 × 10−6 | 133 | 7.1 × 10−3 | 12 × 10−4 |
| K48-Ub4 (7 μM) | 6.1 × 10−6 | 146 | 6.6 × 10−3 | 9.0 × 10−4 |
Apparent KD corresponds to KD of the interaction between GST-Hpr1 and UBA-Mex67 ± K48-Ub4 without considering the conformational change whose k2 and k−2 are indicated separately. Data correspond to mean of at least two independent experiments.