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. 1976 Jul 1;157(1):189–195. doi: 10.1042/bj1570189

The separation and characterization of the methylumbelliferyl beta-galactosidases of human liver.

P S Cheetham, N E Dance
PMCID: PMC1163830  PMID: 962854

Abstract

1. A previously uncharacterized form of human liver acid beta-galactosidase (EC 3.2.1.23), possibly a dimer of molecular weight 160 000, was resolved by gel filtration. It has the same ability to hydrolyse GM1 ganglioside as the two other acid beta-galactosidase forms. 2. The low-molecular-weight forms of acid beta-galactosidase undergo salt-dependent aggregation. 3. The high-molecular-weight component may consist of the low-molecular-weight forms bound to membrane fragments. It can be converted completely into a mixture of these forms. 4. The neutral beta-galactosidase activity can be resolved into two forms by DEAE-cellulose chromatography. They differ in their response to Cl-ions. 5. A new nomenclature is suggested for the six beta-galactosidases so far found in human liver. 6. The enzymic constituents of the beta-galactosidase bands resolved by electrophoresis were re-examined. The A band contains three components. A two-dimensional electrophoretic procedure for resolving the A band is described. 7. The effect of neuraminidase treatment on the behaviour of beta-galactosidases in various separation systems is examined.

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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. Estimation of molecular size and molecular weights of biological compounds by gel filtration. Methods Biochem Anal. 1970;18:1–53. [PubMed] [Google Scholar]
  2. BRADY R. O., KANFER J., SHAPIRO D. THE METABOLISM OF GLUCOCEREBROSIDES. I. PURIFICATION AND PROPERTIES OF A GLUCOCEREBROSIDE-CLEAVING ENZYME FROM SPLEEN TISSUE. J Biol Chem. 1965 Jan;240:39–43. [PubMed] [Google Scholar]
  3. Beck C., Tappel A. L. Rat-liver lysosomal beta-glucosidase: a membrane enzyme. Biochim Biophys Acta. 1968 Jan 8;151(1):159–164. doi: 10.1016/0005-2744(68)90170-8. [DOI] [PubMed] [Google Scholar]
  4. Butterworth J., Bain A. D., McCrae W. M. Cystic fibrosis and liver -galactosidase and -glucosidase. Clin Chim Acta. 1972 Oct;41:367–373. doi: 10.1016/0009-8981(72)90533-5. [DOI] [PubMed] [Google Scholar]
  5. Ho M. W., Cheetham P., Robinson D. Hydrolysis of GM1-ganglioside by human liver beta-galactosidase isoenzymes. Biochem J. 1973 Oct;136(2):351–359. doi: 10.1042/bj1360351. [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. Hurler's syndrome: deficiency of a specific beta galactosidase isoenzyme. Science. 1969 Aug 8;165(3893):611–613. doi: 10.1126/science.165.3893.611. [DOI] [PubMed] [Google Scholar]
  8. Hultberg B., Ockerman P. A. Properties of 4-methylumbelliferyl-beta-galactosidase activities in human liver. Scand J Clin Lab Invest. 1969 May;23(3):213–217. doi: 10.3109/00365516909077651. [DOI] [PubMed] [Google Scholar]
  9. Hultbery B., Ockerman P. A. Artificial substrates in the assay of acid glycosidases. Clin Chim Acta. 1972 Jun;39(1):49–58. doi: 10.1016/0009-8981(72)90298-7. [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. Li S. C., Wan C. C., Mazzotta M. Y., Li Y. T. Requirement of an activator for the hydrolysis of sphingoglycolipids by glycosidases of human liver. Carbohydr Res. 1974 May;34(1):189–193. doi: 10.1016/s0008-6215(00)80383-3. [DOI] [PubMed] [Google Scholar]
  12. Norden A. G., O'Brien J. S. Ganglioside GM1 beta-galactosidase: studies in human liver and brain. Arch Biochem Biophys. 1973 Nov;159(1):383–392. doi: 10.1016/0003-9861(73)90465-7. [DOI] [PubMed] [Google Scholar]
  13. Norden A. G., Tennant L. L., O'Brien J. S. GM1 ganglioside beta-galactosidase. A. Purification and studies of the enzyme from human liver. J Biol Chem. 1974 Dec 25;249(24):7969–7976. [PubMed] [Google Scholar]
  14. O'Brien J. E., Majewski J. C. Effects of de-icing salt on ground water characteristics. Environ Lett. 1975;8(4):303–313. doi: 10.1080/00139307509437440. [DOI] [PubMed] [Google Scholar]
  15. Ockerman P. A., Hultberg B. Fractionation of j-methylumbelliferyl-beta-galactosidase activities in liver in gargoylism. Scand J Clin Lab Invest. 1968;22(3):199–202. doi: 10.3109/00365516809166488. [DOI] [PubMed] [Google Scholar]
  16. Ockerman P. A. Mannosidosis: isolation of oligosaccharide storage material from brain. J Pediatr. 1969 Sep;75(3):360–365. doi: 10.1016/s0022-3476(69)80259-3. [DOI] [PubMed] [Google Scholar]
  17. Okada S., O'Brien J. S. Generalized gangliosidosis: beta-galactosidase deficiency. Science. 1968 May 31;160(3831):1002–1004. doi: 10.1126/science.160.3831.1002. [DOI] [PubMed] [Google Scholar]
  18. Pinsky L., Miller J., Shanfield B., Watters G., Wolfe L. S. GM1 gangliosidosis in skin fibroblast culture: enzymatic differences between types 1 and 2 and observations on a third variant. Am J Hum Genet. 1974 Sep;26(5):563–577. [PMC free article] [PubMed] [Google Scholar]
  19. Robinson D. Multiple forms of glycosidases in the normal and pathological states. Enzyme. 1974;18(1):114–135. doi: 10.1159/000459418. [DOI] [PubMed] [Google Scholar]
  20. Suzuki Y., Suzuki K. Glycosphingolipid beta-galactosidases. I. Standard assay procedures and characterization by electrofocusing and gel filtration of the enzymes in normal human liver. J Biol Chem. 1974 Apr 10;249(7):2098–2104. [PubMed] [Google Scholar]
  21. Van Hoof F., Hers H. G. The abnormalities of lysosomal enzymes in mucopolysacc- haridoses. Eur J Biochem. 1968 Dec;7(1):34–44. doi: 10.1111/j.1432-1033.1968.tb19570.x. [DOI] [PubMed] [Google Scholar]
  22. Winchester B. G. Comparison of the N-acetylhexosaminidase components in the ram testis and epididymis by isoelectric focusing. Biochem J. 1971 Oct;124(5):929–934. doi: 10.1042/bj1240929. [DOI] [PMC free article] [PubMed] [Google Scholar]

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