Summary
Polyclonal antibodies were obtained from rabbits by injection of iduronate sulfatase purified 35,000-fold from human placenta, after elution of the enzyme from sodium dodecyl sulfate (SDS) polyacrylamide gels. The specificity of these antibodies towards iduronate sulfatase was demonstrated by immunoprecipitation of enzyme activity; the level of other lysosomal hydrolases and sulfatases remained constant. Immunoblot of iduronate sulfatase from various human sources showed that the antibody recognises a polypeptide of mol.wt. 72,000 daltons in placenta and serum, and a form of mol.wt. 60,000 daltons in fibroblasts. No immunoprecipitable peptide was found in urine or in the culture medium of fibroblasts. Polypeptides of the same molecular weight were recognised in serum and in fibroblasts of Hunter patients. The presence of altered proteins in these patients was also shown by competition experiments. The addition of Hunter proteins alters the binding of normal enzyme to the antibody.
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References
Archer IM, Harper PS, Wusteman FS (1982) Multiple forms of iduronate 2-sulphate sulphatase in human tissues and body fluids. Biochim Biophys Acta 708:134–140
Bach G, Eisenberg F Jr, Cantz M, Neufeld EF (1973) The defect in the Hunter syndrome: deficiency of sulfoiduronate sulfatase. Proc Natl Acad Sci USA 70:2134–2138
Bell CE, Sly WS, Brot FE (1977) Human β-glucuronidase deficiency mucopolysaccharidosis. Identification of cross-reactive antigen in cultured fibroblasts of deficient patients by enzyme immunoassay. J Clin Invest 59:97–105
Di Natale P, Daniele A (1985) Iduronate sulfatase from human placenta. Biochim Biophys Acta 839:258–261
Di Natale P, Ronsisvalle L (1981) Identification and partial characterization of two enzyme forms of iduronate sulfatase from human placenta. Biochim Biophys Acta 661:106–111
Glössl J, Lembeck K, Gamse G, Kresse H (1980) Morquio disease type A: absence of material cross-reacting with antibodies against N-acetylgalactosamine-6-sulfate sulfatase. Hum Genet 54:87–91
Hall CW, Liebaers I, Di Natale P, Neufeld EF (1978) Enzymic diagnosis of the genetic mucopolysaccharide storage disorders. Methods Enzymol 50:439–456
Hasilik A, Von Figura K (1983) Processing of lysosomal enzymes in fibroblasts. In: Dingle JT, Dean RT, Sly W (eds) Lysosomes in biology and pathology. Elsevier, New York Amsterdam, pp 3–16
Hoogeveen AT, Graham-Kawashima H, D'Azzo A, Galjaard H (1984) Processing of human β-galactosidase in GM1-gangliosidosis and Morquio B syndrome. J Biol Chem 259:1974–1977
Jackson P, Thompson RJ (1984) The immunodetection of brain proteins blotted onto nitrocellulose from fixed and stained two-dimensional polyacrylamide gels. Electrophoresis 5:35–42
Knowles BB, Howe CC, Aden DP (1980) Human hepato-cellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen. Science 209:497–499
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lissens W, Zenati A, Liebaers I (1984) Polyclonal antibodies against iduronate 2-sulphate sulphatase from human urine. Biochim Biophys Acta 801:365–371
Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193: 265–275
McKusick VA, Neufeld EF (1983) The mucopolysaccharide storage diseases. In: Stanbury JB, Wyngaarden JB, Fredrickson DS, Goldstein JL, Brown WJ (eds) The metabolic basis of inherited disease, 4th edn. McGraw Hill, New York, pp 751–777
Roden L (1980) Structure and metabolism of connective tissue proteoglycans. In: Lennarz WJ (ed) The biochemistry of glycoprotein and proteoglycans. Plenum Press, New York, pp 267–371
Rosenstraus M, Chasin L (1975) Isolation of mammalian cell mutants deficient in glucose-6-phosphate dehydrogenase activity: linkage to hypoxanthine phosphoribosyl transferase. Proc Natl Acad Sci USA 72:493–497
Salvatore D, Bonatti S, Di Natale P (1984) Lysosomal α-N-acetylglucosaminidase: purification and characterization of the human urinary enzyme. Bull Mol Biol Med 9:111–121
Shapira E, De Gregorio RR, Matalon R, Nadler HL (1975) Reduced arylsulfatase B activity of the mutant enzyme protein in Maroteaux-Lamy syndrome. Biochem Biophys Res Commun 62:448–455
Sjoberg I, Fransson LA, Matalon R, Dorfman A (1973) Hunter's syndrome: a deficiency of l-idurono-sulfate sulfatase. Biochem Biophys Res Commun 54:1125–1132
Steckel F, Hasilik A, Von Figura K (1983) Biosynthesis and maturation of arylsulfatase B in normal and mutant cultured human fibroblasts. J Biol Chem 258:14322–14326
Tager JM (1985) Biosynthesis and deficiency of lysosomal enzymes. Trends Biochem Sci 10:324–326
Von Figura K, Hasilik A (1984) Genesis of lysosomal enzyme deficiencies. Trends Biochem Sci 9:29–31
Von Figura K, Kresse H (1976) Sanfilippo disease type B: presence of material cross-reacting with antibodies against α-N-acetylglucosaminidase. Eur J Biochem 61:581–588
Waheed A, Hasilik A, Von Figura K (1982) Enhanced breakdown of arylsulfatase A in multiple sulfatase deficiency. Eur J Biochem 123:317–321
Wasteson A, Neufeld EF (1982) Iduronate sulfatase from human plasma. Methods Enzymol 83:573–578
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Daniele, A., Di Natale, P. Hunter syndrome: presence of material cross-reacting with antibodies against iduronate sulfatase. Hum Genet 75, 234–238 (1987). https://doi.org/10.1007/BF00281065
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DOI: https://doi.org/10.1007/BF00281065