TABLE 1.
Excreted end products of glucose metabolism by procyclic T. brucei cell lines
The spent PBS medium of trypanosome cell lines incubated in the presence of 4 mm [1-13C]glucose, was analyzed by 13C NMR spectrometry to detect and quantify 13C-enriched excreted end products.
| Cell linea | nb |
13C-enriched excreted molecules |
|||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Succinate | Acetate | Lactate | Malate | Fumarate | Alanine | β-hydroxybutyrate | Pyruvate | Glycerol | Total | ||
| nmol/h/mg of protein | |||||||||||
| WT (EATRO1125.T7T)c | 5 | 470 ± 88 | 186 ± 21 | 55.0 ± 15 | 20.0 ± 1.7 | 4.0 ± 1.7 | 8.0 ± 2.1 | NDd | ND | <2e | 743 ± 124 |
| Δppdk | 3 | 500 ± 61 | 141 ± 30 | 45.1 ± 10 | 13.7 ± 6.4 | 2.2 ± 1.1 | 6.2 ± 3.9 | ND | ND | <2 | 708 ± 53 |
| Δpepcke | 3 | 1.2 ± 1.4 | 154 ± 54 | 18.0 ± 3.7 | ND | ND | 14.0 ± 8.7 | 8.5 ± 4.4 | ND | 6.3 ± 4.6 | 208 ± 58 |
| Δppdk/Δpepck | 3 | ND | 40 ± 6.5 | 7.0 ± 1.0 | ND | ND | 9.3 ± 2.6 | ND | ND | 17.4 ± 3.7 | 74 ± 4 |
| Δppdk/Δpepck/PEPCK+.i | 2 | 493 | 168 | 15.8 | 26.6 | 5.0 | 1.4 | ND | ND | <2 | 712 |
| PGKC+.ni | 1 | 353 | 151 | 11.6 | 30.5 | 5.3 | 2.4 | ND | ND | <2 | 554 |
| PGKC+.i (1 day) | 1 | 210 | 141 | 38.5 | 11.5 | 1.5 | 2.4 | 2.9 | 2.0 | <2 | 414 |
| PGKC+.i (2 days) | 3 | 91 ± 31 | 118 ± 33 | 39.8 ± 9.2 | 4.2 ± 3.3 | ND | 3.4 ± 1.2 | 2.2 ± 0.8 | 1.7 ± 3.0 | <2 | 261 ± 68 |
| Δppdk/Δpepck/PGKC+.i (3 days) | 3 | 2.9 ± 0.5 | 181 ± 14 | 108.1 ± 9.6 | ND | ND | 7.8 ± 1.1 | 9.2 ± 1.6 | ND | 16.4 ± 2.0 | 325 ± 28 |
a .i: RNAi cell lines tetracycline-induced during 1 to 3 days depending on the cell line and the experiments; .ni: non-induced RNAi cell lines.
b Number of biological replicates.
c From Ebikeme et al. (22).
d Non detectable.
e Since the glycerol peak is embedded in the acylglycerol massif, the detection limit of glycerol was considered as 2 nmol/h/mg of protein.