Abstract
The segregation of human lysosomal arylsulfatase A (ARS-A) has been evaluated in 50 primary hybrid clones derived from four separate fusions involving WBCs from two unrelated individuals and three hamster cell lines. ARS-A was expressed in the hybrids as a dimeric molecule of very similar or identical subunits. The expression of this enzyme was concordant with that of mitochondrial aconitase (ACON-M), an isozyme assigned to chromosome 22, in all 50 clones and with chromosome 22 segregation in all but one of the 29 karyotyped hybrids. No other human chromosome cosegregated with 22 in these clones, suggesting that this enzyme is specified in hybrid cells by a locus (or loci) on a single chromosome. β-Galactosidase (B-GAL) expression was analyzed with two different electrophoresis systems and with a number of cell extract preparation methods in 39 of the primary hybrid clones. The B-GAL isozyme expressed in these hybrid cells was concordant with the expression of glutathione peroxidase-1 (GPX-1), an isozyme assigned to chromosome 3, in all 39 clones and with the segregation of this chromosome in 97% of the 29 karyotyped hybrids. These observations substantiate the prior tentative assignments of an ARS-A locus to chromosome 22 and a B-GAL locus to chromosome 3 (Bruns et al., 1978a, b). The implications of the chromosome assignments of loci for 12 human lysosomal enzymes for the cellular assembly of these organelles are discussed.
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Bach, G., Eisenberg, F., Cantz, M., and Neufeld, E. F. (1973). The defect in the Hunter syndrome: Deficiency of sulfoiduronate sulfatase. Proc. Natl. Acad. Sci. 702134.
Bach, G., and Suzuki, K. (1975). Heterogeneity of human hepatic N-acetyl-β-d-hexosaminidase A activity toward natural glycosphingolipid substrates. J. Biol. Chem. 2501328.
Baggiolini, M., Hirsch, J. G., and de Duve (1969). Resolution of granules from rabbit heterophil leukocytes into distinct populations by zonal centrifugation. J. Cell Biol. 40529.
Bainton, D. F., Ullyot, J. L., and Farquhar, M. G. (1971). The development of neutrophilic polymorphonuclear leukocytes in human bone marrow. J. Exp. Med. 134907.
Balasubramanian, K. A., and Bachhawat, B. K. (1975). Purification, properties and glycoprotein nature of arylsulfatase A from sheep brain. Biochim. Biophys. Acta 403113.
Baum, H., Dodgson, K. S., and Spencer, B. (1959). The assay of arylsulfatases A and B in human urine. Clin. Chim. Acta 4453.
Beutler, E., and West, C. (1974). Red cell glutathione peroxidase polymorphism in Afro-Americans. Am. J. Hum. Genet. 26255.
Blobel, G., and Dobberstein, B. (1975). Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. J. Cell Biol. 67835.
Breen, G. A. M., Lusis, A. J., and Paigen, K. (1977). Linkage of genetic determinants for mouse β-galactosidase electrophoresis and activity. Genetics 8173.
Bretz, U., and Baggiolini, M. (1974). Biochemical and morphological characterization of azurophil and specific granules of human neutrophilic polymorphonuclear leukocytes. J. Cell Biol. 63251.
Bruns, G. A. P., and Gerald, P. S. (1974). Human acid phosphatase in somatic cell hybrids. Science 184480.
Bruns, G. A. P., Mintz, B. J., Leary, A. C., Regina, V. M., and Gerald, P. S. (1978a). Expression of human arylsulfatase A in man-hamster somatic cell hybrids. In Winnipeg Conference (1977): Fourth International Workshop on Human Gene Mapping, Vol. 14, Birth Defects: Original Article Series, The National Foundation, New York, p. 182.
Bruns, G. A. P., Leary, A. C., Regina, V. M., and Gerald, P. S. (1978b). Expression of lysosomal β-d-galactosidase in man-hamster somatic cell hybrids. In Winnipeg Conference (1977): Fourth International Workshop on Human Gene Mapping, Vol. 14, Birth Defects: Original Article Series, The National Foundation, New York, p. 177.
Caspersson, T., Zech, L., Johansson, C., and Modest, E. J. (1970). Identification of human chromosomes by DNA-binding fluorescent agents. Chromosoma 30215.
Champion, M. J., and Shows, T. B. (1977). Mannosidosis: Assignment of the lysosomal α-mannosidase B gene to chromosome 19 in man. Proc. Natl. Acad. Sci. 742968.
Cheetham, P. S. J., and Dance, N. E. (1976). The separation and characterization of the methylumbelliferyl β-galactosidases of human liver. Biochem. J. 157189.
DeLuca, C., Brown, J.A., and Shows, T.B. (1979). Lysosomal arylsulfatase deficiencies in humans: chromosome assignments for arylsulfatase A and B. Proc. Natl. Acad. Sci. 761957.
De Wit, J., Hoeksema, H.L., Bootsma, D., and Westerveld, A. (1979). Assignment of structural β-galactosidase loci to human chromosomes 3 and 22. Human Genet. 49:In press.
De Wit, J., Hoeksema, H. L., Halley, D., Hagemeijer, A., Bootsma, D., and Westerveld, A. (1977). Regional localization of a β-galactosidase locus on human chromosome 22. Somat. Cell Genet. 3351.
de Wit-Verbeek, H. A., Hoogeveen, A., and Galjaard, H. (1978). Complementation studies with enucleated fibroblasts from different variants of β-galactosidase deficiency. Exp. Cell Res. 113215.
Draper, R. K., Fiskum, G. M., and Edmond, J. (1976). Purification, molecular weight, amino acid and subunit composition of arylsulfatase A from human liver. Arch. Biochem. Biophys. 177525.
Flohé, L., Eisele, B., and Wendel, A. (1971). Gluthion-peroxidase. I. Reindarstellung und molekulargewichtsbestimmungen. Hoppe-Seyler's Z. Physiol. Chem. 352151.
Friend, K. K., Chen, S., and Ruddle, F. H. (1976). Differential staining of interspecific chromosomes in somatic cell hybrids by alkaline Giemsa stain. Somat. Cell Genet. 2183.
Giblett, E. R. (1976). Report of the committee on nomenclature. In Baltimore Conference (1975): Third International Workshop on Human Gene Mapping. Vol. 12. Birth Defects: Original Article Series. The National Foundation, New York, p. 65.
Gilbert, F., Kucherlapati, R., Creagan, R. P., Murnane, M. J., Darlington, G. J., and Ruddle, F. H. (1975). Tay-Sach's and Sandhoff's diseases: The assignment of genes for hexosaminidase A and B to individual human chromosomes. Proc. Natl. Acad. Sci. 72263.
Gillin, F. D., Roufa, O. J., Beaudet, A. L., and Caskey, C. T. (1972). 8-Azaguanine resistance in mammalian cells. I. Hypoxanthine-guanine phosphoribosyl-transferase. Genetics 72239.
Grzeschik, K. H. (1976). Assignment of a structural gene for β-glucuronidase to human chromosome C7. Somat. Cell Genet. 2401.
Grzeschik, K. H., Grzeschik, A. M., Banhof, S., Romeo, G., Siniscalco, M., van Someren, H., Meera Khan, P., Westerveld, A., and Bootsma, D. (1972). X-linkage of human α-galactosidase. Nature New Biol. 24048.
Harinath, B. C., and Robins, E. (1971). Arylsulfatases in human brain: Separation, purification and certain properties of the two soluble arylsulfatases. J. Neurochem. 18245.
Hedrick, J. L., and Smith, A. J. (1968). Size and charge isomer separation and estimation of molecular weight of proteins by disc gel electrophoresis. Arch. Biochem. Biophys. 126155.
Hellkuhl, B., and Grzeschik, K. H. (1978). Assignment of a gene for arylsulfatase B (E.C. 3.1.6.1) using human-mouse somatic cell hybrids. In Winnipeg Conference (1977): Fourth International Workshop on Human Gene Mapping, Vol. 14, Birth Defects: Original Article Series, The National Foundation, New York, p. 203.
Ho, M. W., and O'Brien, J. S. (1971). Differential effect of chloride ions on β-galactosidase isozymes: A method of separate assay. Clin. Chim. Acta 32443.
Ingram, P. H., Bruns, G. A. P., Regina, V. M., Eisenman, R. E., and Gerald, P. S. (1977). Expression of α-d-mannosidase in man-hamster somatic cell hybrids. Biochem. Genet. 15455.
Lalley, P. A., Rattazzi, M. C., and Shows, T. B. (1974). Human β-d-N-acetylhexosaminidase A and B: Expression and linkage relationships in somatic cell hybrids. Proc. Natl. Acad. Sci. 711569.
Lalley, P. A., Brown, J. A., Eddy, R. L., Haley, L. L., and Shows, T. B. (1977). Human β-glucuronidase: Assignment of the structural gene to chromosome 7 using somatic cell hybrids. Biochem. Genet. 15367.
Latt, S. A., Willard, H. F., and Gerald, P. S. (1976). BrdU-33258 Hoechst analysis of DNA replication in human lymphocytes with supernumerary or structurally abnormal X chromosomes. Chromosoma 57135.
Lee, G. D., and Van Etten, R. L. (1975). Purification and properties of a homogeneous arylsulfatase A from rabbit liver. Arch. Biochem. Biophys. 166280.
Lowry, O. H., Rosenbrough, N. J., Farr, A. L., and Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193265.
Lusis, A. J., Breen, G. A. M., and Paigen, K. (1977). Nongenetic heterogeneity of mouse β-galactosidase. J. Biol. Chem. 2524613.
McKusick, V. A., and Ruddle, F. H. (1977). The status of the gene map of the human chromosomes. Science 196390.
Mehl, E., and Jatzkewitz, H. (1968). Cerebroside 3-sulfate as a physiological substrate of arylsulfatase. A. Biochim. Biophys. Acta 151619.
Meera Khan, P., Wijnen, L. M. M., and Pearson, P. L. (1978). Assignment of the mitochondrial aconitase gene (ACON M ) to human chromosome 22. In Winnipeg Conference (1977): Fourth International Workshop on Human Gene Mapping, Vol. 14, Birth Defects: Original Article Series, The National Foundation, New York, p. 212.
Neuwelt, E., Stumpf, D., Austin, J., and Kohler, P. (1971). A monospecific antibody to human arylsulfatase A: Preparation, characterization and significance. Biochim. Biophys. Acta 236333.
Nja, A. (1946). A sex-linked type of gargoylism. Acta Paediatr. 33267.
Norden, A. G. W., and O'Brien, J. S. (1973). Ganglioside GM1 β-galactosidase: Studies in human liver and brain. Arch. Biochem. Biophys. 159383.
Norden, A. G. W., Tennant, L. L., and O'Brien, J. S. (1974). GM1 ganglioside β-galactosidase A. J. Biol. Chem. 2497969.
O'Brien, J. S. (1976). Ganglioside storage disease: Recent advances. Tr. Am. Neurol. Assoc. 101102.
O'Brien, J. F., Cantz, M., and Spranger, J. (1974). Maroteaux-Lamy disease (mucopolysaccharidosis VI), subtype A: Deficiency of N-acetylgalactosamine-4-sulfatase. Biochem. Biophys. Res. Commun. 601170.
Porter, M. T., Fluharty, A. L., and Kihara, H., (1969). Metachromatic leukodystrophy: Arylsulfatase-A deficiency in skin fibroblast cultures. Proc. Natl. Acad. Sci. 62887.
Porter, M. T., Fluharty, A. L., Trammel, J., and Kihara, H. (1971). A correlation of intracellular cerebroside sulfatase activity in fibroblasts with latency in metachromatic leukodystrophy. Biochem. Biophys. Res. Commun. 44660.
Rattazzi, M. C., Marks, J. S., and Davidson, R. G. (1973). Electrophoresis of arylsulfatase from normal individuals and patients with metachromatic leukodystrophy. Am. J. Hum. Genet. 25310.
Reuser, A. J. J., Andria, G., de Wit-Verbeek, E., Hoogeveen, E., and Halley, D. (1979). A two-year-old patient with an atypical expression of GM1-β-galactosidase deficiency: Biochemical, immunological and cell genetic studies. Hum. Genet. 4611.
Rosenstraus, M., and Chasin, L. A. (1975). Isolation of mammalian cell mutants deficient in glucose-6-phosphate dehydrogenase activity: Linkage to hypoxanthine-guanine phosphoribosyl transferase. Proc. Natl. Acad. Sci. 72493.
Rushton, A. R., and Dawson, G. (1977) Genetic linkage studies of the human glycosphingolipid β-galactosidases. Biochem. Genet. 151071.
Shows, T. B., Scrafford-Wolff, L., Brown, J. A., and Meisler, M. (1978). Assignment of a β-galactosidase gene (βGAL A ) to chromosome 3 in man. In Winnipeg Conference (1977): Fourth International Workshop on Human Gene Mapping, Vol. 14, Birth Defects: Original Article Series. The National Foundation, New York, p. 219.
Shows, T. B., Scrafford-Wolff, L., Brown, J. A., and Meisler, M. (1979). GM1-gangliosidosis: Chromosome 3 assignment of the β-galactosidase-A gene (βGAL A ). Somat. Cell Genet. 5147.
Slaughter, C. A., Hopkinson, D. A., and Harris, H. (1975). Aconitase polymorphism in man. Ann. Hum. Genet. 39193.
Slaughter, C. A., Povey, S., Carritt, B., Solomon, E., and Bobrow, M. (1978). Assignment of the locus ACON M to chromosome 22. In Winnipeg Conference (1977): Fourth International Workshop on Human Gene Mapping, Vol. 14, Birth Defects: Original Article Series, The National Foundation, New York, p. 223.
Solomon, E., Bobrow, M., Goodfellow, P. N., Bodmer, W. F., Swallow, D. M., Povey, S., and Noel, B. (1976). Human gene mapping using a X/autosome translocation. Somat. Cell Genet. 2125.
Solomon, E., Swallow, D., Burgess, S., and Evans, L. (1979). Assignment of the human acid α-glucosidase gene (αGLU) to chromosome 17 using somatic cell hybrids. Ann. Hum. Genet. 42273.
Sparkes, R. S., Mohandas, T., Sparkes, M. C., and Shulkin, J. D. (1978). Aconitase (E,C, 4.2.1.3) mitochondrial locus mapped to human chromosome 22: Studies with Chinese hamster-human somatic cell hybrids. Biochem. Genet. 16751.
Stevens, R. L., Fluharty, A. L., Skokut, M. H., and Kihara, H. (1975). Purification and properties of arylsulfatase A from human urine. J. Biol. Chem. 2502495.
Stevens, R. L., Fluharty, A. L., Killgrove, A. R., and Kihara, H. (1976). Microheterogeneity of arylsulfatase A from human tissues. Biochim. Biophys. Acta 445661.
Tanaka, H., and Suzuki, K. (1975). Lactosylceramide β-galactosidase in human sphingolipidoses. J. Biol. Chem. 2502324.
Tomino, H., and Meisler, M. (1975). Biochemical and immunological studies of purified mouse β-galactosidase. J. Biol. Chem. 2507752.
Turner, V. S., Turner, B. M., Kucherlapati, R., Ruddle, F. H., and Hirschhorn, K. (1978). Assignment of the gene locus for human α-l-fucosidase to chromosome 1 by analysis of somatic cell hybrids. Somat. Cell Genet. 445.
van Someren, H. (1974). Segregation of human enzymes and antigen markers in man-Chinese hamster somatic cell hybrids. Thesis, University of Leiden, Leiden, Holland.
Wenger, D. A., Tarby, T. J., and Wharton, C. (1978). Macular cherry-red spots and myoclonus with dementia: Coexistent neuraminidase and β-galactosidase deficiencies. Biochem. Biophys. Res. Commun. 82589.
Westerveld, A., Visser, R. P. L. S., Meera Khan, P., and Bootsma, D. (1971). Loss of human genetic markers in man-Chinese hamster somatic cell hybrids. Nature New Biol. 23420.
Wijnen, L. M. M., Monteba-van Heuvel, M., Pearson, P. L., and Meera Khan, P. (1978). Assignment of a gene for glutathione peroxidase (GPX-1) to human chromosome 3. In Winnipeg Conference (1977): Fourth International Workshop on Human Gene Mapping, Vol. 14. Birth Defects: Original Article Series, The National Foundation, New York, p. 232.
Williams, D. E., and Reisfeld, R. A. (1964). Disc electrophoresis in polyacrylamide gels: Extension to new conditions of pH and buffer. Ann. N. Y. Acad. Sci. 121373.
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This work was supported by NIH Grants HD06285 and HD04807.
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Bruns, G.A.P., Mintz, B.J., Leary, A.C. et al. Human lysosomal genes: Arylsulfatase A and β-Galactosidase. Biochem Genet 17, 1031–1059 (1979). https://doi.org/10.1007/BF00504344
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DOI: https://doi.org/10.1007/BF00504344