Carbon dioxide abolishes the reverse Pasteur effect in Leishmania major promastigotes.
The products released by Leishmania major promastigotes incubated with [1-13C]glucose as sole exogenous carbon source were identified using nuclear magnetic resonance (NMR). Under aerobic (95% O2/5% CO2) conditions, acetate, succinate, and small amounts of pyruvate, D-lactate, and glycerol were released in addition to CO2. Under anaerobic (95% N2/5% CO2) conditions, the relative amounts of products formed changed and alanine was also released. The changes in the rates of glucose consumption and product formation during the aerobic to anaerobic transition were measured. Under hypoxic conditions (O2 less than 0.2%), glucose consumption was decreased by about 50%. Under completely anaerobic conditions (100% N2), glucose consumption almost ceased (a total reverse Pasteur effect). The inclusion of 5% CO2 in the gas phase restored hypoxic and anaerobic glucose consumption to the aerobic rate, and increased production of succinate, pyruvate, and D-lactate. Thus, CO2 and very low concentrations of O2 have strong regulatory effects on L. major glucose metabolism. A quantitative carbon balance showed that the NMR-identified products accounted for only about 25% of the glucose carbons consumed under aerobic conditions. CO2, measured as the release of 14CO2 from [U-14C]glucose, accounted for an additional 25% of the glucose consumed. About 11% of the glucose carbon was incorporated into trichloroacetic acid-insoluble products, mostly lipid. Large amounts of label from [U-14C]glucose were incorporated into the intracellular pools of alanine, glutamate, glutamine, and aspartate, indicating that CO2 from unlabeled amino acids contributed to the carbon balance. Under anaerobic conditions, all the glucose carbons consumed could be accounted for solely by the NMR-identified products.
Darling
T N
TN
Department of Cell Biology, Duke University Medical Center, Durham, NC 27710.
Davis
D G
DG
London
R E
RE
Blum
J J
JJ
D000596:Amino Acids / Q000378:metabolism; D000693:Anaerobiosis; D000818:Animals; D002245:Carbon Dioxide / Q000378:metabolism*; D006019:Glycolysis; D007895:Leishmania tropica / Q000378:metabolism*; D008055:Lipids / Q000096:biosynthesis; D009682:Magnetic Resonance Spectroscopy; D009584:Nitrogen / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D010313:Partial Pressure; D013386:Succinates / Q000096:biosynthesis; D019802:Succinic Acid
D000596:Amino Acids; D008055:Lipids; D013386:Succinates; D002245:Carbon Dioxide; D019802:Succinic Acid; D009584:Nitrogen; D010100:Oxygen