Skip to main content
PMC home page
American Journal of Human Genetics logoLink to American Journal of Human Genetics
. 1981 Nov;33(6):916–927.

Feline mucopolysaccharidosis VI: purification and characterization of the resident arylsulfatase B activity.

D T Vine, M M McGovern, M E Haskins, R J Desnick
PMCID: PMC1685156  PMID: 6798861

Abstract

Hepatic arylsulfatase B (ASB) from normal and mucopolysaccharidosis VI (MPS VI) cats was purified over 2,800- and 1,800-fold, respectively, and their physical and kinetic properties were characterized. In contrast to the normal feline enzyme, the partially purified MPS VI residual activity had a 100-fold greater Km value and was markedly less stable to thermal, cryo-, and pH-inactivation. In addition, the MPS VI enzyme had a more negative charge as determined by its migration on polyacrylamide gel electrophoresis and its elution profile on cation exchange chromatography. Finally, the MPS VI activity had approximately half the apparent molecular weight of the normal feline enzyme, which was a homodimer, suggesting that the genetic mutation in feline MPS VI altered the subunit association as well as the kinetic and stability properties of the mutant protein.

Full text

PDF
927

Images in this article

919Fig. 1
on p.919
921Fig. 3
on p.921
925Fig. 9
on p.925

Selected References

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

  1. Agogbua S. I., Wynn C. H. Purification and properties of arylsulphatase B of human liver. Biochem J. 1976 Feb 1;153(2):415–421. doi: 10.1042/bj1530415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andrews P. Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochem J. 1964 May;91(2):222–233. doi: 10.1042/bj0910222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BAUM H., DODGSON K. S., SPENCER B. The assay of arylsulphatases A and B in human urine. Clin Chim Acta. 1959 May;4(3):453–455. doi: 10.1016/0009-8981(59)90119-6. [DOI] [PubMed] [Google Scholar]
  4. Balasubramanian K. A., Bachhawat B. K. Partial purification, properties and glycoproteins nature of arylsulphatase B from sheep brain. J Neurochem. 1976 Aug;27(2):485–492. doi: 10.1111/j.1471-4159.1976.tb12272.x. [DOI] [PubMed] [Google Scholar]
  5. Beratis N. G., Turner B. M., Weiss R., Hirschhorn K. Arylsulfatase B deficiency in Maroteaux-Lamy syndrome: Cellular studies and carrier identification. Pediatr Res. 1975 May;9(5):475–480. doi: 10.1203/00006450-197505000-00003. [DOI] [PubMed] [Google Scholar]
  6. Blakesley R. W., Boezi J. A. A new staining technique for proteins in polyacrylamide gels using coomassie brilliant blue G250. Anal Biochem. 1977 Oct;82(2):580–582. doi: 10.1016/0003-2697(77)90197-x. [DOI] [PubMed] [Google Scholar]
  7. Bleszynski W. S., Roy A. B. Some properties of the sulphatase B of ox brain. Biochim Biophys Acta. 1973 Jul 12;317(1):164–171. doi: 10.1016/0005-2795(73)90209-2. [DOI] [PubMed] [Google Scholar]
  8. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  9. Desnick R. J., Sharp H. L., Grabowski G. A., Brunning R. D., Quie P. G., Sung J. H., Gorlin R. J., Ikonne J. U. Mannosidosis: clinical, morphologic, immunologic, and biochemical studies. Pediatr Res. 1976 Dec;10(12):985–996. doi: 10.1203/00006450-197612000-00008. [DOI] [PubMed] [Google Scholar]
  10. Farooqui A. A., Roy A. B. The sulphatase of ox liver. XX. The preparation of sulphatases B1alpha and B1beta. Biochim Biophys Acta. 1976 Dec 8;452(2):431–439. doi: 10.1016/0005-2744(76)90193-5. [DOI] [PubMed] [Google Scholar]
  11. Fisher R. A., Turner B. M., Dorkin H. L., Harris H. Studies on human erythrocyte inorganic pyrophosphatase. Ann Hum Genet. 1974 Jan;37(3):341–353. doi: 10.1111/j.1469-1809.1974.tb01841.x. [DOI] [PubMed] [Google Scholar]
  12. Fluharty A. L., Edmond J. Arylsulfatases A and B from human liver. Methods Enzymol. 1978;50:537–547. doi: 10.1016/0076-6879(78)50060-8. [DOI] [PubMed] [Google Scholar]
  13. Fluharty A. L., Stevens R. L., Sanders D. L., Kihara H. Arylsulfatase B deficiency in Maroteaux-Lamy syndrome cultured fibroblasts. Biochem Biophys Res Commun. 1974 Jul 24;59(2):455–461. doi: 10.1016/s0006-291x(74)80001-x. [DOI] [PubMed] [Google Scholar]
  14. Haskins M. E., Jezyk P. F., Patterson D. F. Mucopolysaccharide storage disease in three families of cats with arylsulfatase B deficiency: leukocyte studies and carrier identification. Pediatr Res. 1979 Nov;13(11):1203–1210. doi: 10.1203/00006450-197911000-00001. [DOI] [PubMed] [Google Scholar]
  15. Hedrick J. L., Smith A. J. Size and charge isomer separation and estimation of molecular weights of proteins by disc gel electrophoresis. Arch Biochem Biophys. 1968 Jul;126(1):155–164. doi: 10.1016/0003-9861(68)90569-9. [DOI] [PubMed] [Google Scholar]
  16. Jezyk P. F., Haskins M. E., Patterson D. F., Mellman W. J., Greenstein M. Mucopolysaccharidosis in a cat with arylsulfatase B deficiency: a model of Maroteaux-Lamy syndrome. Science. 1977 Nov 25;198(4319):834–836. doi: 10.1126/science.144321. [DOI] [PubMed] [Google Scholar]
  17. Okada K., Collins J., Worth W. S., Austin J. H. Studies of human sulfatase "B." I. Purification and properties from liver. Biochem Med. 1975 Mar;12(3):290–301. doi: 10.1016/0006-2944(75)90131-3. [DOI] [PubMed] [Google Scholar]
  18. REISFELD R. A., LEWIS U. J., WILLIAMS D. E. Disk electrophoresis of basic proteins and peptides on polyacrylamide gels. Nature. 1962 Jul 21;195:281–283. doi: 10.1038/195281a0. [DOI] [PubMed] [Google Scholar]
  19. ROY A. B. The sulphatase of ox liver. I. The complex nature of the enzyme. Biochem J. 1953 Jan;53(1):12–15. doi: 10.1042/bj0530012. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rattazzi M. C., Marks J. S., Davidson R. G. Electrophoresis of arylsulfatase from normal individuals and patients with metachromatic leukodystrophy. Am J Hum Genet. 1973 May;25(3):310–316. [PMC free article] [PubMed] [Google Scholar]
  21. Shafit-Zagardo B., Devine E. A., Desnick R. J. Electrophoretic separation of neutral and acid beta-glucosidase isozymes in human tissues. Biochim Biophys Acta. 1980 Aug 7;614(2):459–465. doi: 10.1016/0005-2744(80)90235-1. [DOI] [PubMed] [Google Scholar]
  22. Shapira E., DeGregorio R. R., Matalon R., Nadler H. L. Reduced arylsulfatase B activity of the mutant enzyme protein in Maroteaux-Lamy syndrome. Biochem Biophys Res Commun. 1975 Jan 20;62(2):448–455. doi: 10.1016/s0006-291x(75)80159-8. [DOI] [PubMed] [Google Scholar]
  23. Shapira E., Nadler H. L. Purification and some properties of soluble human liver arylsulfatases. Arch Biochem Biophys. 1975 Sep;170(1):179–187. doi: 10.1016/0003-9861(75)90109-5. [DOI] [PubMed] [Google Scholar]
  24. Stumpf D. A., Austin J. H., Crocker A. C., LaFrance M. Mucopolysaccharidosis type VI (Maroteaux-Lamy syndrome). I. Sulfatase B deficiency in tissues. Am J Dis Child. 1973 Dec;126(6):747–755. doi: 10.1001/archpedi.1973.02110190597003. [DOI] [PubMed] [Google Scholar]

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

RESOURCES