Table 1 Effects of CYP5122A1 expression on the sterol profiles of L. donovani promastigotes at the log and stationary phases

From: CYP5122A1 encodes an essential sterol C4-methyl oxidase in Leishmania donovani and determines the antileishmanial activity of antifungal azoles

 

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

  1. 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.
  2. aEpisterol, fecosterol and 4-methylzymosterol could not be chromatographically separated.
  3. bZymosterol and Cholesta-7,24-dienol could not be chromatographically separated.
  4. ND not detected. Accumulated: This sterol was detected in the genetic mutants but not in Ld1S WT.
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