The reactions of the phosphorylated pathway of L-serine biosynthesis: thermodynamic relationships in rabbit liver in vivo.
The thermodynamic relationships among the reactions of the phosphorylated pathway of L-serine biosynthesis have been determined in rabbit liver in vivo in different dietary states. The mass action ratios of the reactions involved were calculated from the concentrations of appropriate metabolites in freeze-clamped liver and compared with the equilibrium constants of the same reactions previously determined under physiological conditions. Toward this goal, a new, highly specific enzymatic assay for L-phosphoserine was developed to allow the accurate measurement of this intermediate in biological material. The level of L-phosphoserine, the immediate precursor of L-serine, varied significantly with diet, being 0.81, 0.38, and 0.21 mumol/g wet wt in the fed, and 24 h and 48 h fasted states, respectively. The tissue content of L-phosphoserine was also sensitive to anoxia, falling almost fivefold within 5 min after the liver was removed. Values of for the combined reactions of the first two steps of the pathway of L-serine biosynthesis [D-3-phosphoglycerate dehydrogenase (EC 1.1.1.95) and L-phosphoserine aminotransferase (EC 2.6.1.52)] in livers from animals in different dietary states were calculated to be 1.2 X 10(-4) (fed), 1.4 X 10(-4) (24 h starved), and 0.70 X 10(-4) (48 h starved), all being very close to the value of the combined equilibrium constant of the same reactions (2.44 X 10(-4). Even when there were major changes in the individual components of, such as a fivefold drop in L-phosphoserine and a sevenfold fall in alpha-ketoglutarate following 5 min of anoxia, remained relatively unchanged (2.7 X 10(-4). Thus, it has been concluded that, in rabbit liver under most normal conditions, the combined reactions of D-3-phosphoglycerate dehydrogenase and L-phosphoserine aminotransferase remain very near equilibrium, and that almost all of the disequilibrium of the pathway, amounting to a delta G of -5.5 kcal/mol in the fed state, is at the last step, the L-phosphoserine phosphatase reaction (EC 3.1.3.3).
Lund
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K
Merrill
D K
DK
Guynn
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RW
D000818:Animals; D002237:Carbohydrate Dehydrogenases / Q000378:metabolism; D005215:Fasting; D008099:Liver / Q000201:enzymology / Q000378:metabolism*; D008297:Male; D010084:Oxidation-Reduction; D010100:Oxygen / Q000494:pharmacology; D050543:Phosphoglycerate Dehydrogenase; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D010766:Phosphorylation; D010768:Phosphoserine / Q000378:metabolism*; D011817:Rabbits; D012694:Serine / Q000031:analogs & derivatives* / Q000096:biosynthesis*; D013816:Thermodynamics; D000637:Transaminases / Q000378:metabolism
D010768:Phosphoserine; D012694:Serine; D002237:Carbohydrate Dehydrogenases; D050543:Phosphoglycerate Dehydrogenase; D000637:Transaminases; C022393:phosphoserine aminotransferase; D010744:Phosphoric Monoester Hydrolases; C022394:phosphoserine phosphatase; D010100:Oxygen