Parallel determination of enzyme activities and in vivo fluxes in Brassica napus embryos grown on organic or inorganic nitrogen source.
After the completion of the genomic sequencing of model organisms, numerous post-genomic studies, integrating transcriptome and metabolome data, are aimed at developing a more complete understanding of cell physiology. Here, we measure in vivo metabolic fluxes by steady state labeling, and in parallel, the activity of enzymes in central metabolism in cultured developing embryos of Brassica napus. Embryos were grown on either the amino acids glutamine and alanine as an organic nitrogen source, or on ammonium nitrate as an inorganic nitrogen source. The type of nitrogen made available to developing embryos caused substantial differences in fluxes associated with the tricarboxylic acid cycle, including flux reversion. The changes observed in enzyme activity were not consistent with our estimates of metabolic flux. Furthermore, most extractable enzyme activities are in large surplus relative to the requirements for the observed in vivo fluxes. The results demonstrate that in this model system the metabolic response of central metabolism to changes in environmental conditions can be achieved largely without regulatory reprogramming of the enzyme machinery.
Junker
Björn H
BH
Biology Department, Brookhaven National Laboratory, Upton, NY 11973, United States.
Lonien
Joachim
J
Heady
Lindsey E
LE
Rogers
Alistair
A
Schwender
Jörg
J
D000409:Alanine / Q000378:metabolism; D000596:Amino Acids / Q000032:analysis / Q000378:metabolism; D029688:Brassica napus / Q000196:embryology* / Q000201:enzymology; D002952:Citric Acid Cycle; D046508:Culture Techniques; D005227:Fatty Acids / Q000032:analysis / Q000378:metabolism; D005947:Glucose / Q000032:analysis / Q000378:metabolism; D005973:Glutamine / Q000378:metabolism; D008954:Models, Biological; D009566:Nitrates / Q000378:metabolism; D009584:Nitrogen / Q000378:metabolism*; D012639:Seeds / Q000201:enzymology* / Q000254:growth & development
D000596:Amino Acids; D005227:Fatty Acids; D009566:Nitrates; D005973:Glutamine; D005947:Glucose; D009584:Nitrogen; D000409:Alanine; C006568:ammonium nitrate