Some physical and chemical properties of crystalline alpha-glycerophosphate dehydrogenase.
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Pizer, Lewis I. (University of Pennsylvania, Philadelphia), and Mary L. Potochny. Nutritional and regulatory aspects of serine metabolism in Escherichia coli. J. Bacteriol. 88:611-619. 1964.-Growth studies with a serine auxotroph have demonstrated a relationship between the serine supplied and the extent of growth, which closely agrees with the value calculated from the reported chemical composition of Escherichia coli. Serine could be replaced by glycine and, to a limited extent, by l-threonine. Both exogenous serine and glycine regulate their own endogenous synthesis from glucose or fructose. The inhibition of endogenous synthesis of both amino acids by serine was greater than 90% with both carbon sources. Greater amounts of exogenous glycine were utilized for cell synthesis when fructose was the carbon source. Growth conditions affected the levels of phosphoglycerate dehydrogenase found in cell-free extracts. The highest levels were found in glucose-grown cells and the lowest in cells grown in a medium augmented with l-threonine, l-methionine, l-leucine, and dl-isoleucine. The levels of serine phosphate phosphatase were not altered by changes in growth conditions.
1. Activities of several enzymes of glycolysis, glycerolphosphate cycle, TCA-cycle, beta-oxidation and pentose phosphate cycle were determined in pyloric caeca and ovaries of Asterias rubens during the reproductive cycle. 2. In the ovaries activities of CS and LDH were significantly decreased during ovarian stages 3 and 4, and those of GDH and GAPDH during stage 3 and 4, respectively. 3. In the pyloric caeca activities of GAPDH and LDH were lower in stage 4 and that of GDH in stage 3, whereas those of G6PDH and HOAD were increased in stage 3. 4. Changes observed in enzyme activities are discussed in relation to the reproductive cycle and to the multiple function of the pyloric caeca.
Genetic analysis of backcross progeny from previously characterized rat inbred strains revealed that the biochemical marker Lap-1 is localized in linkage group I (LG I). Lap-1 codes for leucine arylaminopeptidase (EC 3.4.11). The distances of Lap-1 to c, RT6, and Hbb, based on recombination frequencies, are 3.1 +/- 1.5, 8.3 +/- 4.0, and 11.4 +/- 2.8 cM, respectively. Acon-1 codes for aconitase (EC 4.2.1.3). The calculated distances of Acon-1 to c and Hbb are 30.1 +/- 5.0 and 36.1 +/- 5.3 cM, respectively. This suggests that Acon-1 is also in LG I, but the observed high frequency of double crossovers requires further confirmation of this linkage. Ahd-2, Es-6, and Gdc-1 are linked neither to markers of LG I nor to one another.
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Steady-state and transient-state kinetic experiments have been performed to test the proposal that there is a direct (channelled) transfer of NADH from one dehydrogenase to another if the two enzymes exhibit distinct chiral coenzyme specificity (A-side vs. B-side). The results lend no support to this proposal, but are fully consistent with a free-diffusion mechanism of NADH transfer irrespective of the chiral specificity of the enzymes.
Lipogenesis and insulin sensitivity are evaluated in adipose tissue, liver, and diaphragm of ob/ob and non-ob/ob mice. In ob/ob mice, hepatic fatty acid synthesis from [U-14C]glucose is elevated by 4 wk of age, and adipose tissue fatty acid synthesis increases at approximately 7 wk. Hepatic activities in ob/ob mice of glucose-6-phosphate dehydrogenase (EC 1.1.1.49), 6-phosphogluconate dehydrogenase (EC 1.1.1.44), malate dehydrogenase (EC 1.1.1.40), and alpha-glycerophosphate dehydrogenase (EC 1.1.1.8) are dramatically increased by 7 wk of age. Diminished insulin-stimulated glycogen synthesis is first noted in the diaphragm of ob/ob mice at 7 wk of age. Insulin-stimulated glycogen synthesis in adipose tissue of ob/ob mice is impaired at 3 wk. At 7 wk, insulin-stimulated fatty acid synthesis in adipose tissue of ob/ob mice is markedly increased. Adipose tissue glyceride-glycerol synthesis continues to increase throughout development, whereas fatty acid synthesis decreases after 7 wk. The data suggest that alterations in lipid synthesis occur very early in the development of ob/ob mouse, prior to expression to overt obesity, at which time a major contribution to lipogenesis is made by the liver. The altered de novo lipogenesis does not precede the reported diminution in energy metabolism.
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The stereospecificity of the enzyme-dependent oxidation of alpha-[4R-2H]NADH has been determined for four dehydrogenases: two pro-R specific enzymes, pig heart malate dehydrogenase and yeast alcohol dehydrogenase; and two pro-S specific enzymes, rabbit muscle glycerol-3-phosphate dehydrogenase and Rhodopseudomonas spheroides 3-hydroxybutyrate dehydrogenase. In all cases, an enzyme-dependent and substrate-specific oxidation to alpha-NAD+ is observed with the stereochemistry of oxidation identical with that found for the oxidation of the correspondingly labeled beta-NADH. The ability of dehydrogenases from diverse sources to utilize alpha-NADH in a stereochemically competent fashion is discussed in relation to proposed interactions between the nicotinamide sugar moiety and active site residues or obligatory alignments of the pyridine and sugar moieties.
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