title: Expression of lysosomal enzymes in human mutant fibroblast-chick erythrocyte heterokaryons.
abstract: The generation of enzymes located in lysosomes, in cytosol or in endoplasmatic reticulum/Golgi complex is studied in heterokaryons in which chick erythrocyte nuclei are reactivated. The lysosomal enzymes, alpha-glucosidase (alpha-glu) and beta-galactosidase (beta-gal), are synthesized in heterokaryons obtained after fusion of chick erythrocytes with human fibroblasts of patients with Pompe's disease (alpha-glu-deficient) and GM1-gangliosidosis (beta-gal-deficient), respectively. The enzymes appear to be of chick origin and their activities can be detected at first around 4 days after fusion, i.e., at a time when the nucleoli in the erythrocyte nuclei have been reactivated. Maximal activities are reached around 15 days after fusion. No generation of the lysosomal enzyme beta-hexosaminidase is detected in the heterokaryons up to 23 days after fusion of chick erythrocyte with either beta-hexosaminidase A- and B-deficient fibroblasts (Sandhoff's disease) or beta-hexosaminidase A-deficient fibroblasts (Tay-Sachs disease). Similarly no expression of the cytosol enzyme glucose-6-phosphate dehydrogenase (G6PD) is fond up to 30 days after fusion, when chick erythrocytes are fused with fibroblasts from two different G6PD-deficient cell strains (residual activities of 4 and 20% respectively). Indirectly we examined N-acetyl-glucosamine-1-phosphate transferase activity, an enzyme located in the endoplasmic reticulum/Golgi region. This enzyme is needed for the phosphorylation of the lysosomal hydrolases and absence of its activity is the cause of the multiple lysosomal enzyme deficiencies in patients with I-cell disease. The retention of both, chick and human beta-galactosidase in the experiments in which I-cell fibroblasts were fused with chick erythrocytes indicates a reactivation of the gene coding for this phosphorylating enzyme. It also implies that this step in the processing of human lysosomal enzymes is not species-specific.
mesh_terms: D002467:Cell Nucleus / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism*; D005347:Fibroblasts; D005696:Galactosidases / Q000096:biosynthesis; D005954:Glucosephosphate Dehydrogenase / Q000096:biosynthesis; D005959:Glucosidases / Q000096:biosynthesis; D006596:Hexosaminidases / Q000096:biosynthesis; D006822:Hybrid Cells; D008247:Lysosomes / Q000201:enzymology*; D010770:Phosphotransferases / Q000096:biosynthesis*; D017855:Transferases (Other Substituted Phosphate Groups)
chemical_list: D005954:Glucosephosphate Dehydrogenase; D010770:Phosphotransferases; D017855:Transferases (Other Substituted Phosphate Groups); C023222:UDPacetylglucosamine-dolichyl-phosphate acetylglucosamine-1-phosphate transferase; D005696:Galactosidases; D005959:Glucosidases; D006596:Hexosaminidases