A natural-abundance 13C-NMR study of Dictyostelium discoideum metabolism. Monitoring of the spore germination process.

Amoebae and spores of the cellular slime mold Dictyostelium discoideum have been investigated by natural-abundance proton-decoupled 13C-NMR spectroscopy. Axenically grown vegetative amoebae have been found to contain, as prominent metabolites, the polyamines 1,3-diaminopropane (3.2 mM), putrescine (9.4 mM) and spermidine (1.7 mM). We also detected lactic acid (4.4 mM) and the following amino acids as free metabolites in concentrations ranging over 1-3 mM: glycine, alanine, glutamine and glutamate. The glycogen level is highly dependent upon growth state, being below the level of NMR detection in early-exponential cells and reaching about 110 mM glucose equivalents in plateau-phase cells. Dormant spores contained high amounts of trehalose (50 mM), glutamine (73 mM) and glutamate (20 mM). The latter two compounds were not reported previously to be present in such high concentrations in Dictyostelium spores. Germination induced by heat-shock activation was monitored by 13C NMR. No change in the major components occurred during the activation step. The progressive disappearance of trehalose during germination correlated with the decrease of glutamine and glutamate. In general, the data suggest that germinated spores contain a composition of free metabolites very similar to that of starved vegetative amoebae.