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. 1978 Aug;75(8):3970–3973. doi: 10.1073/pnas.75.8.3970

Gaucher disease: isolation and comparison of normal and mutant glucocerebrosidase from human spleen tissue.

P G Pentchev, R O Brady, H E Blair, D E Britton, S H Sorrell
PMCID: PMC392911  PMID: 29293

Abstract

Glucocerebrosidase was purified 26,000-fold from spleens from normal humans and from patients with Gaucher disease (Gaucher spleens). The specific activities of the purified normal and mutant enzymes with glucocerebroside as substrate were 8.5 X 10(5) and 5.4 X 10(4) nmol/mg of protein per hr, respectively. The ratio of enzymatic activities was constant throughout the isolation procedure. The two enzymes appeared to be similar by other parameters such as substrate affinity, heat lability, and pH optimum. Immunotitration with glucocerebrosidase antiserum showed equivalent quantities of crossreacting material in extracts of normal and Gaucher spleens. These data strongly suggest that the genetic basis of Gaucher disease is a strucutral mutation of glucocerebrosidase. The results of sodium dodecyl sulfate gel electrophporesis also indicate that there are differences between the normal and the Gaucher disease enzyme.

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Selected References

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

  1. BRADY R. O., KANFER J. N., SHAPIRO D. METABOLISM OF GLUCOCEREBROSIDES. II. EVIDENCE OF AN ENZYMATIC DEFICIENCY IN GAUCHER'S DISEASE. Biochem Biophys Res Commun. 1965 Jan 18;18:221–225. doi: 10.1016/0006-291x(65)90743-6. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Furbish F. S., Blair H. E., Shiloach J., Pentchev P. G., Brady R. O. Enzyme replacement therapy in Gaucher's disease: large-scale purification of glucocerebrosidase suitable for human administration. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3560–3563. doi: 10.1073/pnas.74.8.3560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Goldstone A., Koenig H. Autolysis of glycoproteins in rat kidney lysosomes in vitro. Effects on the isoelectric focusing behaviour of glycoproteins, arylsulphatase and beta-glucuronidase. Biochem J. 1974 Aug;141(2):527–535. doi: 10.1042/bj1410527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Klibansky C., Hoffman J., Zaizov R., Matoth Y., Pinkhas J., de Vries A. Chronic Gaucher's disease: heat-resistance of leukocyte glucocerebrosidase in relation to some clinical parameters. Biomedicine. 1973 Aug 10;19(8):345–348. [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Mella B., Lang D. J. Leukocyte mitosis: suppression in vitro associated with acute infectious hepatitis. Science. 1967 Jan 6;155(3758):80–81. doi: 10.1126/science.155.3758.80. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Pentchev P. G., Brady R. O., Hibbert S. R., Gal A. E., Shapiro D. Isolation and characterization of glucocerebrosidase from human placental tissue. J Biol Chem. 1973 Aug 10;248(15):5256–5261. [PubMed] [Google Scholar]
  12. Pentchev P. G., Brady R. O. The effect of a heat-stable factor in human spleen on glucocerebrosidase and acid -glucosidase activities. Biochim Biophys Acta. 1973 Feb 28;297(2):491–496. doi: 10.1016/0304-4165(73)90096-2. [DOI] [PubMed] [Google Scholar]
  13. Peters S. P., Coyle P., Coffee C. J., Glew R. H. Purification and properties of a heat-stable glucocerebrosidase activating factor from control and Gaucher spleen. J Biol Chem. 1977 Jan 25;252(2):563–573. [PubMed] [Google Scholar]
  14. Schneider E. L., Ellis W. G., Brady R. O., McCulloch J. R., Epstein C. J. Infantile (type II) Gaucher's disease: in utero diagnosis and fetal pathology. J Pediatr. 1972 Dec;81(6):1134–1139. doi: 10.1016/s0022-3476(72)80245-2. [DOI] [PubMed] [Google Scholar]

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