1. D-amino-acid oxidase is involved in D-serine-induced nephrotoxicity
Masao Maekawa, Tadashi Okamura, Noriyuki Kasai, Yuuichi Hori, Karl H Summer, Ryuichi Konno Chem Res Toxicol. 2005 Nov;18(11):1678-82. doi: 10.1021/tx0500326.
D-serine is nephrotoxic in rats. Based on circumstantial evidence, it has been suspected that D-amino-acid oxidase is involved in this nephrotoxicity. Since we found that LEA/SENDAI rats lacked D-amino-acid oxidase, we examined whether this enzyme was associated with D-serine-induced nephrotoxicity using the LEA/SENDAI rats and control F344 rats. When d-propargylglycine, which is known to have a nephrotoxic effect through its metabolism by D-amino-acid oxidase, was injected intraperitoneally into the F344 rats, it caused glucosuria and polyuria. However, injection of d-propargylglycine into LEA/SENDAI rats did not cause any glucosuria or polyuria, indicating that D-amino-acid oxidase is definitely not functional in these rats. D-serine was then injected into the F344 and LEA/SENDAI rats. It caused glucosuria and polyuria in the F344 rats but not in the LEA/SENDAI rats. These results indicate clearly that D-amino-acid oxidase is responsible for the D-serine-induced nephrotoxicity.
2. Nephrotoxicity of D-proparglyglycine in mice
R Konno, M Ikeda, K Yamaguchi, Y Ueda, A Niwa Arch Toxicol. 2000 Oct;74(8):473-9. doi: 10.1007/s002040000156.
When D-propargylglycine was injected intraperitoneally into mice, polyuria, glycosuria, and aminoaciduria were observed as has been previously reported in rats. The urine of the mice treated with D-propargylglycine contained twice as much protein as that of the control mice. Polyacrylamide gel electrophoresis showed a new protein of approximately 62 kDa in the urine of the D-propargylglycine-treated mice. Protein sequencing revealed that this protein was serum albumin. Since the above-mentioned symptoms suggested dysfunction of the renal proximal tubules, the activity of urinary N-acetyl-beta-D-glucosaminidase, a marker enzyme of injury to the proximal tubules, was measured. The urinary enzyme activity was 2.6 times higher in the D-propargylglycine-treated mice than in the control mice. Light- and electron-microscopy showed degenerative and necrotic cells in the straight part of the proximal tubules of the treated mice. However, none of these symptoms was observed in D-propargylglycine-treated mutant mice, lacking D-amino-acid oxidase. These results indicate that D-propargylglycine itself is not nephrotoxic but its metabolite produced by the D-amino-acid oxidase reaction is nephrotoxic and injures proximal tubular cells, resulting in an impairment of the reabsorption of water, glucose, amino acids, and proteins.
3. Properties of D-amino acid oxidase covalently modified upon its oxidation of D-propargylglycine
P Marcotte, C Walsh Biochemistry. 1978 Jul 11;17(14):2864-8. doi: 10.1021/bi00607a026.
Upon oxidation of D-propargylglycine by D-amino acid oxidase, the enzyme is converted by covalent alkylation to catalytic species with different properties from those of native enzyme. At least five distinct modified enzyme species are present in the preparation, as determined by gel electro-focusing. Individual characterization of the components has not yet been attempted. The combined kinetic and spectral properties of the preparation have been studied. The modified enzymes have a marked preference for hydrophobic amino acids: the rates of oxidation decrease in the series D-phenylalanine, D-methionine, D-norleucine, D-norvaline, D-alpha-aminobutyrate, D-alanine. In addition, the observed Kms of the amino acids are increased, especially those of the smaller substrates (D-alanine and D-alpha-aminobutyrate). A primary kinetic isotope effect is observed upon oxidation of amino acids by the modified enzymes, evidence that this catalysis exhibits a different rate-determining step from catalysis by native enzyme. The modified apoenzyme exhibits intense absorbance at 318--320 nm, not present in native enzyme. This chromophore can be partially (75%) removed by treatment of the modified enzyme with hydrazine. However, the activity of native enzyme is not substantially restored by this process, suggesting the existence of superficial alkylations in addition to the modification responsible for the observed changes in kinetic parameters.