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Kinetic parameters of Ub-AMC cleavage catalyzed by USP5 variants.
| USP5 variant | Km (nM) | kcat (min−1) | kcat/ Km (min−1μM−1) |
|---|---|---|---|
| Wild-type (1–835) | 690 ± 3 | 47 ± 1 | 68.1 |
| Minimal catalytic domain (319–835) | n.d. | n.d. | - |
| ΔnUBP (173–835) | n.d. | n.d. | - |
| ΔcUBP (1–835, Δ172–286) | 940 ± 30 | 5.0 ± 0.1 | 5.3 |
| nUBP double mutant (1–835, F32S/F34S) | 1630 ± 100 | 0.20 ± 0.01 | 0.12 |
n.d., not detectable under assay conditions (see Materials and Methods).
