title: beta-Glucosidase isoenzymes in Epstein-Barr virus-transformed lymphoid cell lines from normal subjects and patients with type 1 Gaucher disease.
abstract: Lymphoid cell lines (LCL) from 3 adult patients with non-neuropathic Gaucher disease were established by Epstein-Barr virus (EBV) transformation and were investigated from the view of enzymology. Glucosylceramide-beta-glucosidase (GlcCer-beta-glucosidase) was present in soluble and particulate fraction of LCL from normal subjects and was deficient in type 1 Gaucher LCL; the deficiency of all molecular forms, shown by electrofocusing, indicates that they are coded by the same gene. The existence of two non-specific beta-glucosidases, one soluble (minor), the other membrane-bound (major), was demonstrated in leucocytes and LCL from normals; in Gaucher LCL, these were also present in a normal range. Characteristic properties of the non-specific membrane-bound beta-glucosidase were defined: lability at acidic pH and strong inhibitory effect by detergents. These properties allowed to discriminate it from the lysosomal GlcCer-beta-glucosidase and to define optimal assay conditions for determination of residual GlcCer-beta-glucosidase activity in Gaucher disease, using artificial substrate, without interference of non-specific membrane-bound beta-glucosidase. These results demonstrate that EBV-transformed LCL represent an accurate model system for enzymatic studies of Gaucher disease.
mesh_terms: D000328:Adult; D002460:Cell Line; D002472:Cell Transformation, Viral; D003902:Detergents; D005776:Gaucher Disease / Q000145:classification / Q000201:enzymology*; D005959:Glucosidases / Q000032:analysis* / Q000172:deficiency*; D005962:Glucosylceramidase / Q000032:analysis / Q000172:deficiency*; D004854:Herpesvirus 4, Human; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000032:analysis*; D008214:Lymphocytes / Q000201:enzymology*; D008247:Lysosomes / Q000201:enzymology; D008565:Membrane Proteins / Q000032:analysis; D013379:Substrate Specificity; D001617:beta-Glucosidase / Q000032:analysis*
chemical_list: D003902:Detergents; D007527:Isoenzymes; D008565:Membrane Proteins; D005959:Glucosidases; D001617:beta-Glucosidase; D005962:Glucosylceramidase