Skip to main content
PMC home page
American Journal of Human Genetics logoLink to American Journal of Human Genetics
. 1980 Mar;32(2):158–173.

Leukocyte β-glucosidase in homozygotes and heterozygotes for Gaucher disease

S S Raghavan, J Topol, E H Kolodny
PMCID: PMC1686022  PMID: 6770675

Abstract

Human leukocytes contain at least two isozymes of 4-methylumbelliferyl-β-glucosidase acting optimally at pH 4.0 and 4.8; in Gaucher disease, only the former is deficient. Brief exposure of the leukocyte homogenate to pH 4.0 at room temperature results in irreversible inactivation of the pH 4.8 activity, while the activity at pH 4.0 remains unaffected. The more acidic isozyme is stimulated four- to fivefold by 0.2% sodium taurodeoxycholate (TDC) with a shift in the pH optimum to 5.0. The less acidic isozyme is completely suppressed in the presence of this detergent. Both leukocyte isozymes appear to be membrane-bound since gel filtration of Sephadex G-200 produces only one peak of activity located at the void volume, unlike in liver and kidney where a second peak also can be demonstrated. Heat inactivation analysis indicated that in controls, assayed in the absence of detergent, pH 4.0 activity is more thermostable than pH 4.8 activity. However, in Gaucher disease, the residual β-glucosidase at pH 4.0 is just as thermolabile as the unaffected pH 4.8 activity. Heat inactivation of the enzyme in the presence of TDC resulted in rapid loss of activity, suggesting a direct effect of the bile salt on the configuration of the enzyme decreasing its thermal stability. In the absence of detergent, acid β-glucosidase shows two Km's, one at 3.2 mM and another at 0.9 mM. In the presence of detergent, only the higher Km at 3.3 mM is obtained. In patients with Gaucher disease and in obligate carriers, the Km remains essentially unaffected while the Vmax shows the expected deficiency.

A reliable and reproducible selective assay technique has been developed for the diagnosis of Gaucher disease homozygotes and obligate heterozygotes and for the carrier screening of individuals at risk for this inherited disorder. The efficacy of this technique has been demonstrated by studying the activity in 42 controls, 26 patients, 32 obligate heterozygotes, and 23 healthy relatives of patients with Gaucher disease.

Full text

PDF
158

Selected References

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

  1. Ben-Yoseph Y., Nadler H. L. Pitfalls in the use of artificial substrates for the diagnosis of Gaucher's disease. J Clin Pathol. 1978 Nov;31(11):1091–1093. doi: 10.1136/jcp.31.11.1091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beutler E., Kuhl W. Detection of the defect of Gaucher's disease and its carrier state in peripheral-blood leucocytes. Lancet. 1970 Mar 21;1(7647):612–613. doi: 10.1016/s0140-6736(70)91646-6. [DOI] [PubMed] [Google Scholar]
  3. Beutler E., Kuhl W., Matsumoto F., Pangalis G. Acid hydrolases in leukocytes and platelets of normal subjects and in patients with Gaucher's and Fabry's disease. J Exp Med. 1976 Apr 1;143(4):975–980. doi: 10.1084/jem.143.4.975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Beutler E., Kuhl W. The diagnosis of the adult type of Gaucher's disease and its carrier state by demonstration of deficiency of beta-glucosidase activity in peripheral blood leukocytes. J Lab Clin Med. 1970 Nov;76(5):747–755. [PubMed] [Google Scholar]
  5. Beutler E., Kuhl W., Trinidad F., Teplitz R., Nadler H. Beta-glucosidase activity in fibroblasts from homozygotes and heterozygotes for Gaucher's disease. Am J Hum Genet. 1971 Jan;23(1):62–66. [PMC free article] [PubMed] [Google Scholar]
  6. Brady R. O., Kanfer J. N., Bradley R. M., Shapiro D. Demonstration of a deficiency of glucocerebroside-cleaving enzyme in Gaucher's disease. J Clin Invest. 1966 Jul;45(7):1112–1115. doi: 10.1172/JCI105417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Broadhead D. M., Butterworth J. The diagnosis of Gaucher's disease in liver using 4-methylumbelliferyl-beta-D-glucopyranoside. Clin Chim Acta. 1977 Feb 15;75(1):155–161. doi: 10.1016/0009-8981(77)90512-5. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Butterworth J., Broadhead D. M. Acid beta-glucosidase and the diagnosis of Gaucher's disease in liver and spleen. Clin Chim Acta. 1978 Aug 1;87(3):433–440. doi: 10.1016/0009-8981(78)90189-4. [DOI] [PubMed] [Google Scholar]
  10. Christomanou H., Cáp C., Sandhoff K. Isoelectric focusing pattern of acid hydrolases in cultured fibroblasts, leucocytes and cell-free amniotic fluid. Neuropadiatrie. 1977 Aug;8(3):238–252. doi: 10.1055/s-0028-1091520. [DOI] [PubMed] [Google Scholar]
  11. Ho M. W. Identity of 'acid' beta-glucosidase and glucocerebrosidase in human spleen. Biochem J. 1973 Nov;136(3):721–729. doi: 10.1042/bj1360721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Ho M. W., Seck J., Schmidt D., Veath M. L., Johnson W., Brady R. O., O'Brien J. S. Adult Gaucher's disease: kindred studies and demonstration of a deficiency of acid beta-glucosidase in cultured fibroblasts. Am J Hum Genet. 1972 Jan;24(1):37–45. [PMC free article] [PubMed] [Google Scholar]
  14. Hultberg B., Ockerman P. A. Beta-glucosidase activities in human tissues. Findings in Gaucher's disease. Clin Chim Acta. 1970 Apr;28(1):169–174. doi: 10.1016/0009-8981(70)90176-2. [DOI] [PubMed] [Google Scholar]
  15. Hultberg B., Sjöblad S., Ockerman P. A. 4-Methylumbelliferyl-beta-glucosidase in cultured human fibroblasts from controls and patients with Gaucher's disease. Clin Chim Acta. 1973 Nov 23;49(1):93–97. doi: 10.1016/0009-8981(73)90347-1. [DOI] [PubMed] [Google Scholar]
  16. Kampine J. P., Brady R. O., Kanfer J. N., Feld M., Shapiro D. Diagnosis of gaucher's disease and niemann-pick disease with small samples of venous blood. Science. 1967 Jan 6;155(3758):86–88. doi: 10.1126/science.155.3758.86. [DOI] [PubMed] [Google Scholar]
  17. Kanfer J. N., Raghavan S. S., Mumford R. A. Hydrolytic and transglucosylation activities of a purified calf spleen beta-glucosidase. Biochim Biophys Acta. 1975 May 23;391(1):129–140. doi: 10.1016/0005-2744(75)90159-x. [DOI] [PubMed] [Google Scholar]
  18. Klibansky C., Hoffman J., Zaizov R., Matoth Y. Gaucher's disease, chronic adult type: a comparative study of glucocerebroside and methylumbelliferyl-glucopyranoside cleaving potency in leukocytes. Biomedicine. 1974 Jan;20(1):24–30. [PubMed] [Google Scholar]
  19. Kolodny E. H., Mumford R. A. Human leukocyte acid hydrolases: characterization of eleven lysosomal enzymes and study of reaction conditions for their automated analysis. Clin Chim Acta. 1976 Jul 15;70(2):247–257. doi: 10.1016/0009-8981(76)90426-5. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Lisman J. J., Hooghwinkel G. J. Human brain beta-glucosidase and beta-galactosidase. Influence of triton X-100 on the extraction, activation and gel chromatographic behaviour. Neurobiology. 1973;3(4):264–274. [PubMed] [Google Scholar]
  22. Mueller O. T., Rosenberg A. beta-Glucoside hydrolase activity of normal and glucosylceramidotic cultured human skin fibroblasts. J Biol Chem. 1977 Feb 10;252(3):825–829. [PubMed] [Google Scholar]
  23. Ockerman P. A. Identity of beta-glucosidase, beta-xylosidase and one of the beta-galactosidase activities in human liver when assayed with 4-methylumbelliferyl-beta-D-glycosides studies in cases of Gaucher's disease. Biochim Biophys Acta. 1968 Aug 6;165(1):59–62. [PubMed] [Google Scholar]
  24. Owada M., Sakiyama T., Kitagawa T. Neuropathic Gaucher's disease with normal 4-methylumbelliferyl-beta-glucosidase activity in the liver. Pediatr Res. 1977 May;11(5):641–646. doi: 10.1203/00006450-197705000-00004. [DOI] [PubMed] [Google Scholar]
  25. Peters S. P., Coyle P., Glew R. H. Differentiation of beta-glucocerebrosidase from beta-glucosidase in human tissues using sodium taurocholate. Arch Biochem Biophys. 1976 Aug;175(2):569–582. doi: 10.1016/0003-9861(76)90547-6. [DOI] [PubMed] [Google Scholar]
  26. Peters S. P., Lee R. E., Glew R. H. A microassay for Gaucher's disease. Clin Chim Acta. 1975 May 1;60(3):391–396. doi: 10.1016/0009-8981(75)90083-2. [DOI] [PubMed] [Google Scholar]
  27. Turner B. M., Beratis N. G., Hirschhorn K. Cell-specific differences in membrane beta-glucosidase from normal and Gaucher cells. Biochim Biophys Acta. 1977 Feb 9;480(2):442–449. doi: 10.1016/0005-2744(77)90036-5. [DOI] [PubMed] [Google Scholar]
  28. Wenger D. A. Assay of beta-glucosidase and sphingomyelinase for identification of patients and carriers of Gaucher's and Niemann-Pick diseases. Adv Exp Med Biol. 1978;101:707–717. doi: 10.1007/978-1-4615-9071-2_65. [DOI] [PubMed] [Google Scholar]
  29. Wenger D. A., Clark C., Sattler M., Wharton C. Synthetic substrate beta-glucosidase activity in leukocytes: a reproducible method for the identification of patients and carriers of Gaucher's disease. Clin Genet. 1978 Feb;13(2):145–153. doi: 10.1111/j.1399-0004.1978.tb04242.x. [DOI] [PubMed] [Google Scholar]

Articles from American Journal of Human Genetics are provided here courtesy of American Society of Human Genetics

RESOURCES