Table II.
Kinetic Analysis of Guinea Pig 11β-HSD1 Wild-Type and Mutant Proteins
| Dehydrogenase |
Reductase |
|||
|---|---|---|---|---|
| Mutant | Km (μM) | kcat (min−1) | Km (μM) | kcat (min−1) |
| Wild Type | 459 ± 0.49 | 1.02 ± 0.04 | 4.09 ± 0.35 | 0.57 ± 0.01 |
| Y266E | 2.86 ± 0.22 | 0.47 ± 0.01 | 3.67 ± 0.58 | 0.20 ± 0.01 |
| I275E | 1.35 ± 0.40 | 0.24 ± 0.02 | 31.1 ± 9.8 | 0.37 ± 0.04 |
| F278E | 5.73 ± 1.13 | 1.12 ± 0.08 | 3.13 ± 0.63 | 0.48 ± 0.02 |
| L279E | 3.23 ± 0.29 | 0.53 ± 0.01 | 5.27 ± 1.33 | 0.41 ± 0.03 |
Assays were carried out as per Materials and Methods. Km values are given in μM with kcat values given in min−1 (mean ± SEM). Results were analyzed using nonlinear regression by VisualEnzymics (Softzymics).