Regulation of carbon flux from amino acids into sugar phosphates in Xenopus embryos.
Xenopus laevis oocytes and embryos are glycogenic cells, metabolizing sugar phosphates into glycogen. These cells have very low pyruvate kinase activity in vivo and, consequently, make little pyruvate and lactate through glycolysis. Nevertheless, oocytes and embryos do contain significant pyruvate and lactate levels. To determine the source of carbon for sugar phosphates and pyruvate, 14C-labeled intermediary metabolites were injected into fertilized eggs and their metabolism examined by thin-layer chromatography. Alanine, pyruvate, and lactate form a pool of carbon that fluxes into sugar phosphates. Cytosolic (nonmitochondrial) aspartate, oxaloacetate, and malate form a pool of carbon which is largely blocked in the short-term from entering the smaller alanine/pyruvate/lactate pool. The data indicate that the major source of carbon for sugar phosphates in fertilized eggs and rapidly cleaving embryos is the alanine/pyruvate/lactate pool. Pyruvate from this pool is converted in the mitochondria to phosphoenolpyruvate, which in turn is metabolized outside the mitochondria to sugar phosphates. A key enzyme in regulating flux from amino acid carbon to pyruvate is malic enzyme. Three malic enzyme isozymes, one soluble and two mitochondrial, were partially isolated and kinetically characterized from total ovarian tissue. Full-grown oocytes and eggs, however, have very low soluble malic enzyme activity, which results in the separation of the cytosolic aspartate/oxaloacetate/malate and alanine/pyruvate/lactate pools.
Dworkin
M B
MB
Ernst-Boehringer-Institut, Vienna, Austria.
Dworkin-Rastl
E
E
D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism; D002244:Carbon / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D004734:Energy Metabolism; D005260:Female; D005971:Glutamates / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism; D007700:Kinetics; D007773:Lactates / Q000378:metabolism; D008291:Malate Dehydrogenase / Q000378:metabolism; D008293:Malates / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D010053:Ovary / Q000201:enzymology; D010063:Ovum / Q000201:enzymology; D011773:Pyruvates / Q000378:metabolism; D013403:Sugar Phosphates / Q000378:metabolism*; D014982:Xenopus laevis / Q000196:embryology*
D000596:Amino Acids; D005971:Glutamates; D007527:Isoenzymes; D007773:Lactates; D008293:Malates; D011773:Pyruvates; D013403:Sugar Phosphates; D001224:Aspartic Acid; D009249:NADP; D002244:Carbon; C030298:malic acid; D008291:Malate Dehydrogenase