Table 1.
Effects of CYP5122A1 expression on the sterol profiles of L. donovani promastigotes at the log and stationary phases
| Sterols | Stationary phase | Log phase | |||||
|---|---|---|---|---|---|---|---|
| Ld22A1 + /− vs. Ld1S WT | Ld22A1 + /− +pXNG4-22A1 vs. Ld1S WT | Ld22A1- +pXNG4-22A1 vs. Ld1S WT | Ld22A1 + /− vs. Ld1S WT | Ld22A1 + /− +pXNG4-22A1 vs. Ld1S WT | Ld22A1- +pXNG4-22A1 vs. Ld1S WT | ||
| 4- and/or 14-methylated sterols | Lanosterol | 9.3 ± 2.1 | 0.20 ± 0.03 | 0.38 ± 0.06 | 23 ± 5 | 0.35 ± 0.09 | 0.39 ± 0.16 |
| 4-Hydroxylanosterol | 3.3 ± 0.2 | 0.28 ± 0.07 | 0.23 ± 0.00 | 14 ± 1 | 0.21 ± 0.03 | 0.12 ± 0.04 | |
| 4,14-Dimethylzymosterol | 2.0 ± 0.3 | 0.47 ± 0.06 | 0.24 ± 0.01 | 3.3 ± 0.8 | 0.20 ± 0.02 | 0.20 ± 0.07 | |
| 14-Methylzymosterol | 0.81 ± 0.04 | 0.64 ± 0.05 | 0.74 ± 0.03 | 0.73 ± 0.01 | 0.52 ± 0.00 | 0.59 ± 0.20 | |
| 14-Methylfecosterol | ND | Accumulated | ND | ND | Accumulated | ND | |
| 4-Methylzymosterola | 1.6 ± 0.1 | 2.3 ± 0.0 | 3.8 ± 0.1 | 1.1 ± 0.0 | 1.7 ± 0.2 | 1.0 ± 0.3 | |
| FF-MAS | ND | ND | ND | ND | ND | ND | |
| T-MAS | ND | ND | ND | ND | ND | ND | |
| 4,14-demethylated sterols | Zymosterolb | 1.2 ± 0.2 | 1.7 ± 0.1 | 0.89 ± 0.07 | 1.2 ± 0.0 | 1.6 ± 0.1 | 0.30 ± 0.11 |
| Fecosterola | 1.6 ± 0.1 | 2.3 ± 0.0 | 3.8 ± 0.1 | 1.1 ± 0.0 | 1.7 ± 0.2 | 1.0 ± 0.3 | |
| Episterola | 1.6 ± 0.1 | 2.3 ± 0.0 | 3.8 ± 0.1 | 1.1 ± 0.0 | 1.7 ± 0.2 | 1.0 ± 0.3 | |
| Cholesta-7,24-dienolb | 1.2 ± 0.2 | 1.7 ± 0.1 | 0.89 ± 0.07 | 1.2 ± 0.0 | 1.6 ± 0.1 | 0.30 ± 0.11 | |
| 5-dehydro sterols | 7-Dehydrodesmosterol | 0.75 ± 0.14 | 0.47 ± 0.07 | 0.14 ± 0.01 | 0.66 ± 0.10 | 0.48 ± 0.02 | 0.044 ± 0.019 |
| 5-Dehydroepisterol | 0.99 ± 0.18 | 0.83 ± 0.10 | 0.84 ± 0.07 | 0.82 ± 0.07 | 0.67 ± 0.03 | 0.30 ± 0.13 | |
| Ergosta-5,7-dienol | 1.2 ± 0.2 | 1.3 ± 0.2 | 2.0 ± 0.1 | 0.79 ± 0.08 | 0.78 ± 0.07 | 0.52 ± 0.23 | |
| Ergostateraenol | 1.0 ± 0.2 | 0.64 ± 0.10 | 1.0 ± 0.1 | 1.1 ± 0.1 | 0.96 ± 0.15 | 0.30 ± 0.10 | |
| Ergosterol | 1.0 ± 0.2 | 1.2 ± 0.1 | 1.8 ± 0.1 | 0.79 ± 0.14 | 1.0 ± 0.1 | 0.53 ± 0.16 | |
| Cholesterol | 1.2 ± 0.2 | 1.2 ± 0.0 | 1.5 ± 0.0 | 0.87 ± 0.07 | 0.89 ± 0.07 | 0.52 ± 0.17 | |
The sterol levels in genetic mutants were normalized to the corresponding sterol levels in Ld1S WT. The values represent the mean ± 1/2 range (n = 2). Source data are provided as a Source Data file.
aEpisterol, fecosterol and 4-methylzymosterol could not be chromatographically separated.
bZymosterol and Cholesta-7,24-dienol could not be chromatographically separated.
ND not detected. Accumulated: This sterol was detected in the genetic mutants but not in Ld1S WT.