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. 1974 Nov;143(2):295–301. doi: 10.1042/bj1430295

Separation and properties of human brain hexosaminidase C

Isobel Braidman 1, Mark Carroll 1, Norman Dance 1, Donald Robinson 1
PMCID: PMC1168384  PMID: 4477952

Abstract

Hexosaminidase C was separated from human brain supernatant by immunoadsorption of the A and B forms on to a column of immobilized antibody followed by preparative starch-block electrophoresis. There were some differences in the properties of hexosaminidase C preparations after each of these stages, shown by comparison of their heat-inactivation characteristics and filtration through Bio-Gel P-200. The C form prepared by both separation steps had properties which differed markedly from those of the A and B isoenzymes; its molecular weight was much larger, greater than 200000, it had optimum activity between pH6 and 7 and could not be successfully eluted from DEAE-cellulose, even with high salt concentrations, or from Sephadex G-200. These results seem to support the proposal that the C form is under a separate genetic control from the others.

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Selected References

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

  1. Andrews P. The gel-filtration behaviour of proteins related to their molecular weights over a wide range. Biochem J. 1965 Sep;96(3):595–606. doi: 10.1042/bj0960595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Carroll M., Dance N., Masson P. K., Robinson D., Winchester B. G. Human mannosidosis--the enzyme defect. Biochem Biophys Res Commun. 1972 Oct 17;49(2):579–583. doi: 10.1016/0006-291x(72)90450-0. [DOI] [PubMed] [Google Scholar]
  3. Carroll M., Robinson D. Immunological properties of N-acetyl-beta-D-glucosaminidase of normal human liver and of GM2-gangliosidosis liver. Biochem J. 1973 Jan;131(1):91–96. doi: 10.1042/bj1310091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Furth A. J., Robinson D. Specificity and multiple forms of beta-galactosidase in the rat. Biochem J. 1965 Oct;97(1):59–66. doi: 10.1042/bj0970059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. HERS H. G. alpha-Glucosidase deficiency in generalized glycogenstorage disease (Pompe's disease). Biochem J. 1963 Jan;86:11–16. doi: 10.1042/bj0860011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ho M. W., O'Brien J. S. Differential effect of chloride ions on -galactosidase isoenzymes: a method for separate assay. Clin Chim Acta. 1971 May;32(3):443–450. doi: 10.1016/0009-8981(71)90446-3. [DOI] [PubMed] [Google Scholar]
  7. Ho M. W., O'Brien J. S. Gaucher's disease: deficiency of 'acid' -glucosidase and reconstitution of enzyme activity in vitro. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2810–2813. doi: 10.1073/pnas.68.11.2810. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hooghwinkel G. J., Veltkamp W. A., Overdijk B., Lisman J. J. Electrophoretic separation of -N-acetylhexosaminidases of human and bovine brain and liver and of Tay-Sachs brain tissue. Hoppe Seylers Z Physiol Chem. 1972 May;353(5):839–841. [PubMed] [Google Scholar]
  9. KUNKEL H. G. Zone electrophoresis. Methods Biochem Anal. 1954;1:141–170. doi: 10.1002/9780470110171.ch6. [DOI] [PubMed] [Google Scholar]
  10. Kint J. A. Antagonistic action of chondroitin sulphate and cetylpyridinium chloride on human liver beta-galactosidase. FEBS Lett. 1973 Oct 1;36(1):53–56. doi: 10.1016/0014-5793(73)80335-7. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Marsh C. A., Gourlay G. C. Evidence for a non-lysosomal alpha-mannosidase in rat liver homogenates. Biochim Biophys Acta. 1971 Apr 14;235(1):142–148. doi: 10.1016/0005-2744(71)90041-6. [DOI] [PubMed] [Google Scholar]
  13. O'Brien J. S., Okada S., Fillerup D. L., Veath M. L., Adornato B., Brenner P. H., Leroy J. G. Tay-Sachs disease: prenatal diagnosis. Science. 1971 Apr 2;172(3978):61–64. doi: 10.1126/science.172.3978.61. [DOI] [PubMed] [Google Scholar]
  14. O'Brien J. S., Okada S., Ho M. W., Fillerup D. L., Veath M. L., Adams K. Ganglioside storage diseases. Fed Proc. 1971 May-Jun;30(3):956–969. [PubMed] [Google Scholar]
  15. Okada S., O'Brien J. S. Tay-Sachs disease: generalized absence of a beta-D-N-acetylhexosaminidase component. Science. 1969 Aug 15;165(3894):698–700. doi: 10.1126/science.165.3894.698. [DOI] [PubMed] [Google Scholar]
  16. Phillips N. C., Robinson D., Winchester B. G., Jolly R. D. Mannosidosis in Angus cattle. The enzymic defect. Biochem J. 1974 Feb;137(2):363–371. doi: 10.1042/bj1370363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Poenaru L., Dreyfus J. C. Electrophoretic study of hexosaminidases. Hexosaminidase C. Clin Chim Acta. 1973 Feb 12;43(3):439–442. doi: 10.1016/0009-8981(73)90486-5. [DOI] [PubMed] [Google Scholar]
  18. Price R. G., Dance N. The demonstration of multiple heat stable forms of N-acetyl- -glucosaminidase in normal human serum. Biochim Biophys Acta. 1972 Jun 22;271(1):145–153. doi: 10.1016/0005-2795(72)90142-0. [DOI] [PubMed] [Google Scholar]
  19. Robinson D., Jordan T. W., Horsburgh T. The N-acetyl- -D-hexosaminidases of calf and human brain. J Neurochem. 1972 Aug;19(8):1975–1985. doi: 10.1111/j.1471-4159.1972.tb01487.x. [DOI] [PubMed] [Google Scholar]
  20. Robinson D., Stirling J. L. N-Acetyl-beta-glucosaminidases in human spleen. Biochem J. 1968 Apr;107(3):321–327. doi: 10.1042/bj1070321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sandhoff K., Harzer K., Wässle W., Jatzkewitz H. Enzyme alterations and lipid storage in three variants of Tay-Sachs disease. J Neurochem. 1971 Dec;18(12):2469–2489. doi: 10.1111/j.1471-4159.1971.tb00204.x. [DOI] [PubMed] [Google Scholar]
  22. Sandhoff K., Wässle W. Anreicherung und Charakterisierung zweier Formen der menschlichen N-acetyl- -D-hexosaminidase. Hoppe Seylers Z Physiol Chem. 1971 Aug;352(8):1119–1133. [PubMed] [Google Scholar]
  23. Steers E., Jr, Cuatrecasas P., Pollard H. B. The purification of beta-galactosidase from Escherichia coli by affinity chromatography. J Biol Chem. 1971 Jan 10;246(1):196–200. [PubMed] [Google Scholar]
  24. Stirling J. L. Separation and characterisation of N-acetyl- -glucosaminidases A and P from maternal serum. Biochim Biophys Acta. 1972 Jun 22;271(1):154–162. doi: 10.1016/0005-2795(72)90143-2. [DOI] [PubMed] [Google Scholar]
  25. Suzuki Y., Suzuki K. Partial deficiency of hexosaminidase component a in juvenile gm2-gangliosidosis. Neurology. 1970 Sep;20(9):848–851. doi: 10.1212/wnl.20.9.848. [DOI] [PubMed] [Google Scholar]

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