Summary
A series of man-Chinese hamster and man-mouse somatic cell hybrids was investigated to study the localization of the genes coding for the human lysosomal enzyme β-galactosidase (EC 3.2.1.23) and for its protective protein. Using a monoclonal antibody, raised against human placental β-galactosidase, it was observed that the structural locus for the β-galactosidase polypeptide is located on chromosome 3. The nature of the involvement of chromosome 22 in the expression of human β-galactosidase was elucidated by metabolic labelling of the hybrids with radioactive amino acids, immunoprecipitation with monoclonal and polyclonal antibodies against β-galactosidase, followed by analysis via gel electrophoresis and fluorography.
The data show that the presence of chromosome 22 coincides with the presence of a 32 kd protein. This polypeptide, the “protective protein” was previously shown to be intimately associated with human β-galactosidase. In addition, the protective protein was found to be essential for the in vivo stability of β-galactosidase by aggregating β-galactosidase monomers into high molecular weight multimes. Both chromosome 3 and 22 are therefore necessary to obtain normal levels og β-galactosidase activity in human cells.
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Barneveld R, Keijzer W, Tegelaers FPW, Ginns EI, Geurs van Kessel A, Brady RO, Barranger JA, Tager JM, Galjaard H, Westerveld A, Reuser AJJ (1983) Assignment of the gene coding for human β-glucocerebrosidase to the region q21–q31 of chromosome 1 using monoclonal antibodies. Hum Genet 64:227–231
Bonner WM, Laskey RA (1974) A film detection method for tritiumlabelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem 46:83–88
Bruns GAP, Leary AC, Regina VM, Gerald PS (1978) Lysosomal β-D-galactosidase in man-hamster somatic cell hybrids. Cytogenet Cell Genet 22:177–181
D'Azzo A, Hoogeveen A, Reuser AJJ, Robinson D, Galjaard H (1982) Molecular defect in combined β-galactosidase and neuraminidase deficiency in man. Proc Natl Acad Sci USA 79:4535–4539
de Wit J, Hoeksema HL, Halley D, Hagemeijer A, Bootsma D, Westerveld A (1977) Regional localization of a β-galactosidase locus on chromosome 22. Somatic Cell Genet 3:351–363
de Wit J, Hoeksema HL, Bootsma D, Westerveld A (1979) Assignment of structural β-galactosidase loci to human chromosomes 3 and 22. Hum Genet 51:259–267
Galjaard H (1980) Genetic metabolic diseases. Elsevier, North-Holland, pp 817, 825
Galjaard H, D'Azzo A, Hoogeveen A, Verheijen F (1984) Combined β-galactosidase-sialidase deficiency in man: genetic defect of a “protective protein”. In: Barranger JA, Brady RO (eds) Molecular basis of lysosomal storage disorders. Academic Press, Orlando
Halley DJJ, Konings A, Hupkes P, Galjaard H (1984) Regional mapping of the human gene for lysosomal α-glucosidase by in situ hybridization. Hum Genet 67:326–328
Hasilik A, Neufeld EF (1980) Biosynthesis of lysosomal enzymes in fibroblasts. Synthesis as precursors of higher molecular weight. J Biol Chem 255:4937–4945
Hoeksema HL, De Wit J, Westerveld A (1980) The genetic defect in the various types of human β-galactosidase deficiency. Hum Genet 53:241–247
Hoogeveen A, D'Azzo A, Brossmer R, Galjaard H (1981) Correlation of combined β-galactosidase/neuraminidase deficiency in human fibroblasts. Biochem Biophys Res Commun 103:292–300
Hoogeveen AT, Verheijen FW, Galjaard H (1983) The relation between human lysosomal β-galactosidase and its protective protein. J Biol Chem 258:12143–12146
Konings A, Hupkes P, Versteeg R, Grosveld G, Reuser A, Galjaard H (1984) Cloning a cDNA for the lysosomal α-glucosidase. Biochem Biophys Res Commun 119:252–258
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Meera Khan P (1971) Enzyme electrophoresis on cellulose acetate gel: Zymogram patterns in man-mouse and man-Chinese hamster somatic cell hybrids. Arch Biochem Biophys 145:470–483
Shows TB, Scrafford-Wolff L, Brown JA, Meister M (1978) Assignment of a β-galactosidase gene (β-Gal A) to chromosome 3 in man. Cytogenet Cell Genet 22:219–222
Verheijen F, Brossmer R, Galjaard H (1982) Purification of acid β-galactosidase and acid neuraminidase from bovine testis: Evidence for an enzyme complex. Biochem Biophys Res Commun 108:868–875
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Sips, H.J., de Wit-Verbeek, H.A., de Wit, J. et al. The chromosomal localization of human β-galactosidase revisited: a locus for β-galactosidase on human chromosome 3 and for its protective protein on human chromosome 22. Hum Genet 69, 340–344 (1985). https://doi.org/10.1007/BF00291653
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DOI: https://doi.org/10.1007/BF00291653