Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1977 Sep 1;165(3):497–502. doi: 10.1042/bj1650497

Purification and some properties of 1-aspartamido-beta-N-acetylglucosamine amidohydrolase from human liver.

B Dugal, J Stromme
PMCID: PMC1164932  PMID: 21658

Abstract

Human liver 1-aspartamido-beta-N-acetylglucosamine amidohydrolase (aspartylglucosylaminase, EC 3.5.1.26) was purified 17 500-fold to apparent homogeneity as judged from polyacrylamide-gel disc electrophoresis. A pH optimum of 7.7-9.0 was found. The Km value was pH- and temperature-dependent. At 37 degrees C and pH 7.7, Km was 0.16 mM and it increased to 0.29 at pH 6.0 and 0.23 at pH 9.0. At 25 degrees C and pH 7.7, a Km value of 0.99 mM was obtained. When the substrate concentration was varied, apparent Michaelis-Menten kinetics were obtained. p-Hydroxymercuribenzoate, glutathione or cysteine had no effect on the enzyme activity; 5 mM-N-acetylcysteine inhibited about 47% of the total enzyme activity. Apart from Cu2+, other bivalent ions were virtually ineffective at 1 mM. The kinetic study differentiates this enzyme from aspartylglucosylaminase from other sources.

Full text

PDF
500

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aula P., Näntö V., Laipio M. L., Autio S. Aspartylglucosaminuria: deficiency of aspartylglucosaminidase in cultured fibroblasts of patients and their heterozygous parents. Clin Genet. 1973;4(3):297–300. doi: 10.1111/j.1399-0004.1973.tb01161.x. [DOI] [PubMed] [Google Scholar]
  2. Aula P., Raivio K., Autio S. Enzymatic diagnosis and carrier detection of aspartylglucosaminuria using blood samples. Pediatr Res. 1976 Jun;10(6):625–629. doi: 10.1203/00006450-197606000-00012. [DOI] [PubMed] [Google Scholar]
  3. Conchie J., Strachan I. Distribution, purification and properties of 1-aspartamido-beta-N-acetylglucosamine amidohydrolase. Biochem J. 1969 Dec;115(4):709–715. doi: 10.1042/bj1150709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dugal B. S. Apparent co-operative effect of hydrogen carbonate (HCO-3) on pigeon kidney pyruvate carboxylase. Enzyme. 1976;21(4):317–326. doi: 10.1159/000458874. [DOI] [PubMed] [Google Scholar]
  5. Dugal B. S. Intracellular localization of pyruvate carboxylase in mammalian liver. Exp Cell Res. 1974 Nov;89(1):63–70. doi: 10.1016/0014-4827(74)90187-6. [DOI] [PubMed] [Google Scholar]
  6. Dugal B. S. Localization, purification and substrate specificity of monoamine oxidase. Biochim Biophys Acta. 1977 Jan 11;480(1):56–69. doi: 10.1016/0005-2744(77)90320-5. [DOI] [PubMed] [Google Scholar]
  7. Dugal B. S., Louis B. Pyruvatkarboxylase aus Taubenleber. Reinigung und Eigenschaften des Enzyms. Acta Biol Med Ger. 1974;32(1):1–16. [PubMed] [Google Scholar]
  8. Dugal B. Measurement of 1-aspartamido-beta-N-acetylglucosamine amidohydrolase activity in human tissues. Biochem J. 1977 Apr 1;163(1):9–14. doi: 10.1042/bj1630009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dugal Birinder S. Apparent co-operative effect of acetyl-CoA on pigeon liver pyruvate carboxylase. FEBS Lett. 1973 Mar 1;30(2):181–184. doi: 10.1016/0014-5793(73)80647-7. [DOI] [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. MURAKAMI M., EYLAR E. H. BETA-(N-ACETYLGLUCOSAMINE)-N-GLYCOSIDASE: AN ENZYME WHICH CATALYZES THE HYDROLYSIS OF 1-BETA-ASPARTYL-2-ACETAMIDO-1,2-DIDEOXY-D-GLUCOSYLAMINE. J Biol Chem. 1965 Jan;240:PC556–PC558. [PubMed] [Google Scholar]
  12. Mahadevan S., Tappel A. L. Beta-aspartylglucosylamine amido hydrolase of rat liver and kidney. J Biol Chem. 1967 Oct 25;242(20):4568–4576. [PubMed] [Google Scholar]
  13. Makino M., Kojima T., Ohgushi T., Yamashina I. Studies on enzymes acting on glycopeptides. J Biochem. 1968 Feb;63(2):186–192. doi: 10.1093/oxfordjournals.jbchem.a128760. [DOI] [PubMed] [Google Scholar]
  14. Makino M., Kojima T., Yamashina I. Enzymatic cleavage of glycopeptides. Biochem Biophys Res Commun. 1966 Sep 22;24(6):961–966. doi: 10.1016/0006-291x(66)90344-5. [DOI] [PubMed] [Google Scholar]
  15. Malik M. N., Stracher A. Allosteric behavior of platelet myosin. Arch Biochem Biophys. 1977 Jan 30;178(2):451–458. doi: 10.1016/0003-9861(77)90215-6. [DOI] [PubMed] [Google Scholar]
  16. Ohgushi T., Yamashina I. Distribution of a glycopeptide-degrading enzyme in tissue and cells. Biochim Biophys Acta. 1968 Mar 11;156(2):417–419. doi: 10.1016/0304-4165(68)90276-6. [DOI] [PubMed] [Google Scholar]
  17. Pollitt R. J., Jenner F. A. Enzymatic cleavage of 2-acetamido-1-(beta'-L-aspartamido)-1,2-dideoxy-beta-D-glucose by human plasma and seminal fluid. Failure to detect the heterozygous state for aspartylglycosaminuria. Clin Chim Acta. 1969 Sep;25(3):413–416. doi: 10.1016/0009-8981(69)90201-0. [DOI] [PubMed] [Google Scholar]
  18. Pollitt R. J., Jenner F. A., Merskey H. Aspartylglycosaminuria. An inborn error of metabolism associated with mental defect. Lancet. 1968 Aug 3;2(7562):253–255. doi: 10.1016/s0140-6736(68)92355-6. [DOI] [PubMed] [Google Scholar]
  19. Savolainen H. Isolation of the liver N-aspartyl-beta-glucosaminidase in aspartylglucosaminuria. Biochem J. 1976 Mar 1;153(3):749–750. doi: 10.1042/bj1530749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tarentino A. L., Maley F. The purification and properties of a beta-aspartyl N-acetylglucosylamine amidohydrolase from hen oviduct. Arch Biochem Biophys. 1969 Mar;130(1):295–303. doi: 10.1016/0003-9861(69)90036-8. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES