Table 2.
Relative activities of wild-type and chimeric KshA5 with a range of steroid substrates
| Steroid substrated | Relative KSH activitye |
||||
|---|---|---|---|---|---|
| A5c | A5A1β | A5A1loop | A5A1β207-216 | A5W248D | |
| 4-Androstene-3,17-dione (AD) | 100 | 100 | 100 | 100 | 100 |
| 1,4-Androstadiene-3,17-dione (ADD) | 51 ± 4 | 189 ± 35a | 114 ± 19a | 53 ± 4 | 66 ± 8a |
| 4-Androstene-17β-ol-3-one (testosterone) | 113 ± 20 | 155 ± 19a | 125 ± 13 | 95 ± 7 | 109 ± 7 |
| 4-Pregnene-3,20-dione (progesterone) | 65 ± 1 | 179 ± 37a | 106 ± 5a | 95 ± 21 | 74 ± 9 |
| 19-Nor-4-androstene-3,17-dione (nordion) | 111 ± 18 | 28 ± 5a | 27 ± 4a | 118 ± 7 | 102 ± 10 |
| 1-(5α)-Androstene-3,17-dione | 99 ± 9 | 28 ± 6a | 33 ± 3a | 94 ± 20 | 94 ± 12 |
| 5α-Androstane-3,17-dione | 73 ± 6 | 13 ± 1a | 14 ± 7a | 69 ± 9 | 58 ± 14 |
| 5β-Androstane-3,17-dione | 99 ± 12 | 20 ± 10a | 25 ± 5a | 50 ± 16a | 55 ± 3a |
| 5-Cholestene-3β-ol (cholesterol)b | — | — | — | — | — |
| 5α-Androstane-17β-ol-3-one (stanolon) | 100 ± 10 | — | 16 ± 3a | 95 ± 8 | 57 ± 19a |
| 3α-Hydroxy-5α-pregnane-20-oneb | — | — | — | — | — |
| 11β-Hydrocortisone | 93 ± 9 | 22 ± 5a | 24 ± 11a | 91 ± 10 | 94 ± 7 |
| 3β-Hydroxy-5α-androstane-17-one | — | — | — | — | — |
| 23,24-Bisnorcholesta-4-ene-22-oic acid (4-BNC) | 88 ± 11 | 216 ± 44a | 120 ± 29 | 89 ± 21 | 50 ± 37 |
| 9α-Hydroxy-4-androstene-3,17-dione (9OHAD) | — | — | — | — | — |
a
P < 0.02 (analysis of variance).
b
Steroid concentration is 25 μM due to low solubility.
c
Data taken from Petrusma et al. (22).
d
For chemical structures of steroids used in this study, see Fig. S1 in the supplemental material.
e
Relative KSH activities (with standard deviations) compared to activity with AD, which is set at 100% (Table 4). Steroid substrates were tested at a concentration of 200 μM. —, no initial KSH activity detectable.