TABLE 2.
Relative initial activities of wild-type and mutated KshA1N with a range of steroid substrates
| Steroid substrate | Relative initial activity of Ksh (%)a |
||||
|---|---|---|---|---|---|
| A1N | A1V207T | A1P210E | A1D225R | A1N238D | |
| 4-Androstene-3,17-dione (AD) | 100 | 100 | 100 | 100 | 100 |
| 4-Androstene-17β-ol-3-one (testosterone) | 135 ± 10 | 161 ± 30 | 154 ± 20 | 133 ± 37 | 139 ± 24 |
| 1,4-Androstadiene-3,17-dione (ADD) | 282 ± 40 | 289 ± 38 | 275 ± 47 | 178 ± 29 | 252 ± 50 |
| 1,4-Androstadiene-17β-ol-3-one (boldenone) | 252 ± 31 | 258 ± 33 | 255 ± 51 | 174 ± 18 | 267 ± 29 |
| 1-(5α)-Androstene-3,17-dione | 91 ± 19 | 112 ± 20 | 100 ± 22 | 63 ± 18 | 95 ± 27 |
| 5α-Androstane-3,17-dione | 23 ± 4 | 32 ± 16 | 32 ± 6 | NDc | 26 ± 9 |
| 5β-Androstane-3,17-dione | 33 ± 6 | 42 ± 18 | 41 ± 12 | 15 ± 5 | 52 ± 19 |
| 5α-Androstane-17β-ol-3-one (androstanolone) | 18 ± 3 | 35 ± 11 | 13 ± 7 | ND | 25 ± 9 |
| 17α-Methyl-1-(5α)-androstane-17β-ol-3-one | 78 ± 8 | 92 ± 21 | 82 ± 10 | 32 ± 13 | 73 ± 32 |
| 4-Androstene-3,17-dione-19-ol | 10 ± 2 | 12 ± 4 | 16 ± 2 | ND | ND |
| 4-Pregnene-3,20-dione (progesterone) | 335 ± 38 | 430 ± 55 | 323 ± 40 | 215 ± 61 | 320 ± 40 |
| 11β-Hydrocortisone | 52 ± 9 | 82 ± 26 | 42 ± 11 | 22 ± 8 | 57 ± 19 |
| 23,24-Bisnorchol-4-ene-22-oic acid (4-BNC) | 426 ± 52 | 499 ± 58 | 447 ± 65 | 260 ± 43 | 440 ± 52 |
| 4-Cholestene-3-oneb (cholestenone) | 21 ± 2 | 56 ± 17 | 30 ± 9 | ND | 25 ± 12 |
a
Relative activity signifies the percentage of activity toward substrates at 200 μM relative to that of AD, which was set at 100%. Errors were calculated as standard errors of the means (from three experiments).
b
Steroid concentration of 25 μM due to the low solubility of cholesterone.
c
ND, no detectable activity toward specific steroid.