Peroxisomal D-amino Acid Oxidase Research Articles

Glycine metabolism in animals and humans: implications for nutrition and health

Glycine is a major amino acid in mammals and other animals. It is synthesized from serine, threonine, choline, and hydroxyproline via inter-organ metabolism involving primarily the liver and kidneys. Under normal feeding conditions, glycine is not adequately synthesized in birds or in other animals, particularly in a diseased state. Glycine degradation occurs through three pathways: the glycine cleavage system (GCS), serine hydroxymethyltransferase, and conversion to glyoxylate by peroxisomal D-amino acid oxidase. Among these pathways, GCS is the major enzyme to initiate glycine degradation to form ammonia and CO2 in animals. In addition, glycine is utilized for the biosynthesis of glutathione, heme, creatine, nucleic acids, and uric acid. Furthermore, glycine is a significant component of bile acids secreted into the lumen of the small intestine that is necessary for the digestion of dietary fat and the absorption of long-chain fatty acids. Glycine plays an important role in metabolic regulation, anti-oxidative reactions, and neurological function. Thus, this nutrient has been used to: (1) prevent tissue injury; (2) enhance anti-oxidative capacity; (3) promote protein synthesis and wound healing; (4) improve immunity; and (5) treat metabolic disorders in obesity, diabetes, cardiovascular disease, ischemia-reperfusion injuries, cancers, and various inflammatory diseases. These multiple beneficial effects of glycine, coupled with its insufficient de novo synthesis, support the notion that it is a conditionally essential and also a functional amino acid for mammals (including pigs and humans).

Why isd-serine nephrotoxic and α-aminoisobutyric acid protective?

D-Serine selectively causes necrosis of S(3) segments of proximal tubules in rats. This leads to aminoaciduria and glucosuria. Coinjection of the nonmetabolizable amino acid alpha-aminoisobutyric acid (AIB) prevents the tubulopathy. D-serine is selectively reabsorbed in S(3), thereby gaining access to peroxisomal D-amino acid oxidase (D-AAO). D-AAO-mediated metabolism produces reactive oxygen species. We determined the fractional excretion of amino acids and glucose in rats after intraperitoneal injection of d-serine alone or together with reduced glutathione (GSH) or AIB. Both compounds prevented the hyperaminoaciduria. We measured GSH concentrations in renal tissue before (control) and after D-serine injection and found that GSH levels decreased to approximately 30% of control. This decrease was prevented when equimolar GSH was coinjected with D-serine. To find out why AIB protected the tubule from D-serine toxicity, we microinfused D-[(14)C]serine or [(14)C]AIB (0.36 mmol/l) together with [(3)H]inulin in late proximal tubules in vivo and measured the radioactivity in the final urine. Fractional reabsorption of D-[(14)C]serine and [(14)C]AIB amounted to 55 and 70%, respectively, and 80 mmol/l of AIB or D-serine mutually prevented reabsorption to a great extent. D-AAO activity measured in vitro (using D-serine as substrate) was not influenced by a 10-fold higher AIB concentration. We conclude from these results that 1) D-AAO-mediated d-serine metabolism lowers renal GSH concentrations and thereby provokes tubular damage because reduction of reactive oxygen species by GSH is diminished and 2) AIB prevents d-serine-induced tubulopathy by inhibition of D-serine uptake in S(3) segments rather than by interfering with intracellular D-AAO-mediated D-serine metabolism.

Differential effect of sunflower seed oil on hepatic and renal d-amino acid oxidase in the rat

1. 1. The specific activity of hepatic and renal peroxisomal d-amino acid oxidase ( d-Aaox) was measured in rats fed diets containing various quantities of vegetable oil. 2. 2. Increasing the amount of dietary sunflower seed oil (SSO) from 10 to 25% (w/w) reduced the specific activity of hepatic d-Aaox by up to 30% after 10 days. 3. 3. In both tissues, the enzyme activity was moderately decreased during the first two-day period after administration of the 25% SSO diet was begun. Unlike hepatic d-Aaox, renal d-Aaox returned to its baseline level in the kidney after the third day. 4. 4. In contrast to SSO, hydrogenated coconut oil (HCO) did not evoke alterations of d-AAOX activity. 5. 5. The activity levels of another peroxisomal enzyme, l-2-hydroxy acid oxidase ( l-Haox), in the liver of rats fed the high-SSO diet vs those fed the control diet were similar. 6. 6. The subcellular distribution of d-Aaox and d-Haox was not altered in the liver of rats fed the 25% SSO diet during the 10-day period.

Inhibition of peroxisomal fatty acyl-CoA oxidase by antimycin A.

Peroxisomal fatty acyl-CoA oxidase was inhibited by micromolar concentrations of antimycin A, an inhibitor of mitochondrial respiration. The inhibition was observed with all three substrates tested, i.e. palmitoyl-CoA, trihydroxycoprostanoyl-CoA and hexadecanedioyl-CoA. The peroxisomal D-amino acid oxidase was also inhibited by antimycin, but the peroxisomal L-alpha-hydroxyacid oxidase and uric acid oxidase and the mitochondrial monoamine oxidase were not. The degree of inhibition of acyl-CoA oxidase by antimycin was strongly dependent on the amount of cellular protein present in the assay mixture: at a fixed antimycin concentration, the inhibition was gradually lost with increasing protein concentrations. At a fixed cellular protein concentration in the assay mixtures, the mitochondrial oxidation of glutamate or palmitoylcarnitine was inhibited at antimycin concentrations that were much lower than those required for the inhibition of fatty acyl-CoA oxidase. Our results, nevertheless, demonstrate that antimycin A must be used with caution, when it is added to homogenates or subcellular fractions in order to distinguish between mitochondrial and peroxisomal fatty acid oxidation.

Pipecolic acid is oxidized by renal and hepatic peroxisomes: Implications for Zellweger's cerebro-hepato-renal syndrome (CHRS)

Increased levels of pipecolic acid have been reported in patients with cerebro-hepatorenal syndrom (CHRS) of Zellweger and the general deficiency of peroxisomal function has been implicated in its pathogenesis. We have therefore investigated the capacity of normal peroxisomes to metabolize pipecolic acid. Highly purified peroxisomes were obtained from rat liver and rat and beef kidney cortex by a recently developed method using metrizamide gradients and a vertical rotor. These preparations oxidized d,l-pipecolic acid as evidenced by the measurement of H 2O 2 production. Incubation with either the d- or l-isomer revealed that almost exclusively d-pipecolate is oxidized. The specific activities proved to be 20–50 times higher in renal than in hepatic peroxisomes. A commercially available crystalline suspension of d-amino acid oxidase from porcine kidney also oxidized the pipecolic acid with the following rates 54:36:1 respectively for d-:, d,l-: l-isomers. Incubation of vibratome sections of rat kidney and liver in a medium containing d-pipecolic acid and cerous ions, revealed electron-dense deposits over the matrix of peroxisomes confirming the localization also by fine structural cytochemistry. These observations demonstrate the capability of mammalian peroxisomes to oxidize pipecolic acid and suggest that the absence or deficiency of peroxisomal d-amino acid oxidase may be implicated in the pathogenesis of hyperpipecolatemia in Zellweger's CHRS.

Interpretation of inverse acclimation to temperature

In kidney of goldfish acclimated to 5, 15 and 25° C the peroxisomal enzyme peroxidase and the peroxisomal and cytoplasmic matrix enzyme catalase showed inverse (Precht type 5) acclimation. Peroxisomal D-amino acid oxidase and lysosomal acid phosphatase were unchanged in activity (Precht type 4). A review of literature data on enzyme acclimation patterns shows that generally enzymes concerned with energy liberation — enzymes of glycolysis, hexose monophosphate shunt, TCA cycle and electron transport, also Na, K-ATPase and the synthetic amino acyl transferase — show compensatory acclimation to temperature (Precht type 3). Enzymes for degradation of metabolic intermediates and products such as peroxisomal and lysosomal enzymes, Mg-ATPase, acetylcholine esterase, show no or inverse acclimation to temperature. Changes in digestive enzymes depend on state of nutrition.