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. 1982 Aug;79(15):4535–4539. doi: 10.1073/pnas.79.15.4535

Molecular defect in combined beta-galactosidase and neuraminidase deficiency in man.

A D'Azzo, A Hoogeveen, A J Reuser, D Robinson, H Galjaard
PMCID: PMC346709  PMID: 6812049

Abstract

In normal human fibroblasts, an enzymically active 85,000-dalton precursor form of beta-galactosidase is processed, via a number of intermediates, into a mature 64,000-dalton form. In addition there is an enzymically inactive 32,000-dalton component and its 54,000-dalton precursor. In fibroblasts from patients with a combined deficiency of beta-galactosidase and neuraminidase these last two components are absent and hardly any mature beta-galactosidase can be demonstrated. Nevertheless, in the mutant fibroblasts, precursor beta-galactosidase is synthesized and processed normally. The excessive intralysosomal degradation that is responsible for the deficiency of mature beta-galactosidase can be partially corrected by addition of the protease inhibitor leupeptin, which results in the accumulation of 85,000-dalton precursor beta-galactosidase and of a partially processed 66,000-dalton form. When mutant cells were grown in the presence of a "corrective factor" purified from the medium of NH4Cl-stimulated cell cultures, both beta-galactosidase and neuraminidase activities were restored to low control levels. The immunoprecipitation pattern was completely normal after addition of the corrective factor, and mature 64,000-dalton beta-galactosidase accumulated in the mutant fibroblasts. We propose that the combined beta-galactosidase/neuraminidase deficiency is caused by a defective 32,000-dalton glycoprotein which is normally required to protect beta-galactosidase and neuraminidase against excessive intralysosomal degradation and to give these enzymes their full hydrolytic activity.

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Selected References

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

  1. Andria G., Del Giudice E., Reuser A. J. Atypical expression of beta-galactosidase deficiency in a child with Hurler-like features but without neurological abnormalities. Clin Genet. 1978 Jul;14(1):16–23. doi: 10.1111/j.1399-0004.1978.tb02055.x. [DOI] [PubMed] [Google Scholar]
  2. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  3. Cantz M., Gehler J., Spranger J. Mucolipidosis I: increased sialic acid content and deficiency of an alpha-N-acetylneuraminidase in cultured fibroblasts. Biochem Biophys Res Commun. 1977 Jan 24;74(2):732–738. doi: 10.1016/0006-291x(77)90363-1. [DOI] [PubMed] [Google Scholar]
  4. Durand P., Gatti R., Cavalieri S., Borrone C., Tondeur M., Michalski J. C., Strecker G. Sialidosis (mucolipidosis I). Helv Paediatr Acta. 1977 Nov;32(4-5):391–400. [PubMed] [Google Scholar]
  5. Frisch A., Neufeld E. F. Limited proteolysis of the beta-hexosaminidase precursor in a cell-free system. J Biol Chem. 1981 Aug 10;256(15):8242–8246. [PubMed] [Google Scholar]
  6. Gonzalez-Noriega A., Grubb J. H., Talkad V., Sly W. S. Chloroquine inhibits lysosomal enzyme pinocytosis and enhances lysosomal enzyme secretion by impairing receptor recycling. J Cell Biol. 1980 Jun;85(3):839–852. doi: 10.1083/jcb.85.3.839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hasilik A., Neufeld E. F. Biosynthesis of lysosomal enzymes in fibroblasts. Synthesis as precursors of higher molecular weight. J Biol Chem. 1980 May 25;255(10):4937–4945. [PubMed] [Google Scholar]
  8. Hoeksema H. L., De Wit J., Westerveld A. The genetic defect in the various types of human beta-galactosidase deficiency. Hum Genet. 1980 Feb;53(2):241–247. doi: 10.1007/BF00273505. [DOI] [PubMed] [Google Scholar]
  9. Hoeksema H. L., van Diggelen O. P., Galjaard H. Intergenic complementation after fusion of fibroblasts from different patients with beta-galactosidase deficiency. Biochim Biophys Acta. 1979 Jan 12;566(1):72–79. doi: 10.1016/0005-2744(79)90250-x. [DOI] [PubMed] [Google Scholar]
  10. Hoogeveen A. T., Verheijen F. W., d'Azzo A., Galjaard H. Genetic heterogeneity in human neuraminidase deficiency. Nature. 1980 Jun 12;285(5765):500–502. doi: 10.1038/285500a0. [DOI] [PubMed] [Google Scholar]
  11. Hoogeveen A., d'Azzo A., Brossmer R., Galjaard H. Correction of combined beta-galactosidase/neuraminidase deficiency in human fibroblasts. Biochem Biophys Res Commun. 1981 Nov 16;103(1):292–300. doi: 10.1016/0006-291x(81)91692-2. [DOI] [PubMed] [Google Scholar]
  12. Kleijer W. J., Hoogeveen A., Verheijen F. W., Niermeijer M. F., Galjaard H., O'Brien J. S., Warner T. G. Prenatal diagnosis of sialidosis with combined neuraminidase and beta-galactosidase deficiency. Clin Genet. 1979 Jul;16(1):60–61. doi: 10.1111/j.1399-0004.1979.tb00851.x. [DOI] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Lowden J. A., O'Brien J. S. Sialidosis: a review of human neuraminidase deficiency. Am J Hum Genet. 1979 Jan;31(1):1–18. [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Skudlarek M. D., Swank R. T. Biosynthesis of two lysosomal enzymes in macrophages. Evidence for a precursor of beta-galactosidase. J Biol Chem. 1979 Oct 25;254(20):9939–9942. [PubMed] [Google Scholar]
  18. Suzuki Y., Nakamura N., Fukuoka K., Shimada Y., Uono M. beta-Galactosidase deficiency in juvenile and adult patients. Report of six Japanese cases and review of literature. Hum Genet. 1977 Apr 15;36(2):219–229. doi: 10.1007/BF00273261. [DOI] [PubMed] [Google Scholar]
  19. Suzuki Y., Sakuraba H., Hayashi K., Suzuki K., Imahori K. Beta-galactosidase-neuraminidase deficiency: restoration of beta-galactosidase activity by protease inhibitors. J Biochem. 1981 Jul;90(1):271–273. doi: 10.1093/oxfordjournals.jbchem.a133462. [DOI] [PubMed] [Google Scholar]
  20. Thomas G. H., Tiller G. E., Jr, Reynolds L. W., Miller C. S., Bace J. W. Increased levels of sialic acid associated with a sialidase deficiency in I-cell disease (mucolipidosis II) fibroblasts. Biochem Biophys Res Commun. 1976 Jul 12;71(1):188–195. doi: 10.1016/0006-291x(76)90267-9. [DOI] [PubMed] [Google Scholar]
  21. Thomas G. H., Tipton R. E., Ch'ien L. T., Reynolds L. W., Miller C. S. Sialidase (alpha-n-acetyl neuraminidase) deficiency: the enzyme defect in an adult with macular cherry-red spots and myoclonus without dementia. Clin Genet. 1978 Apr;13(4):369–379. doi: 10.1111/j.1399-0004.1978.tb01194.x. [DOI] [PubMed] [Google Scholar]
  22. Van Diggelen O. P., Galjaard H., Sinnott M. L., Smith P. J. Specific inactivation of lysosomal glycosidases in living fibroblasts by the corresponding glycosylmethyl-p-nitrophenyltriazenes. Biochem J. 1980 May 15;188(2):337–343. doi: 10.1042/bj1880337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Van Diggelen O. P., Schram A. W., Sinnott M. L., Smith P. J., Robinson D., Galjaard H. Turnover of beta-galactosidase in fibroblasts from patients with genetically different types of beta-galactosidase deficiency. Biochem J. 1981 Oct 15;200(1):143–151. doi: 10.1042/bj2000143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wenger D. A., Tarby T. J., Wharton C. Macular cherry-red spots and myoclonus with dementia: coexistent neuraminidase and beta-galactosidase deficiencies. Biochem Biophys Res Commun. 1978 May 30;82(2):589–595. doi: 10.1016/0006-291x(78)90915-4. [DOI] [PubMed] [Google Scholar]
  25. van Diggelen O. P., Hoogeveen A. T., Smith P. J., Reuser A. J., Galjaard H. Enhanced proteolytic degradation of normal beta-galactosidase in the lysosomal storage disease with combined beta-galactosidase and neuraminidase deficiency. Biochim Biophys Acta. 1982 Apr 21;703(1):69–76. doi: 10.1016/0167-4838(82)90012-7. [DOI] [PubMed] [Google Scholar]

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