Table 2.
lysoGPCho seriesb | |||||
---|---|---|---|---|---|
|
|||||
Cell line | Tet | (−/18:y) | (−/20:y) | (−/22:y) | Total |
WT | 53 | 31 | 72 | 156 | |
Δ pla1 | 15 | 8 | 7 | 30 | |
ovexPLA1-myc | − | 56 | 22 | 81 | 159 |
+ | 159 | 42 | 231 | 432 | |
Δ plal rescPLAI-myc | − | 29 | 14 | 23 | 66 |
+ | 52 | 22 | 54 | 128 |
Total lipid extracts from 108 PCF T. brucei cells spiked with 500 pmol of each internal standard [lysoGPCho(17:0/−) and lysoGPCho(24:0/−)] were examined by ESI-MS-MS. lysoGPCho molecules were detected from precursor ion scanning for m/z 184 (see insets from Fig. 5B), and they were quantified by comparison with the internal standards. Values are from a typical analysis and are presented as pmol per 108 cells. Tet, tetracycline.
Integration of peaks to obtain peak areas was performed by grouping lysoGPCho into various series of peaks whereby one series comprises both the major and minor lysoGPCho molecules, and their isotopes, containing the same FA chain length but with varying degrees of unsaturation, represented by ‘y’.