title: Correlation between enzyme activity and substrate storage in a cell culture model system for Gaucher disease.
abstract: Gaucher disease, the most common sphingolipidosis, is caused by a decreased activity of glucosylceramide beta-glucosidase, resulting in the accumulation of glucosylceramide in macrophage-derived cells known as Gaucher cells. Much of the storage material is thought to originate from the turnover of cell membranes, such as phagocytosed red and white blood cells. In this study, an in vitro model of Gaucher disease was developed by treating the murine macrophage cell line J774 with a specific inhibitor of glucosylceramide beta-glucosidase, conduritol B-epoxide, and feeding red blood cell ghosts, in order to mimic the disease state. It was found in this model system that glucosylceramide beta-glucosidase activity could be reduced to about 11-15% of the normal control level before increased storage of glucosylceramide occurred. This in vitro system allows insight into the correlation between enzyme activity and lipid storage as predicted by the theory of residual enzyme activity that was proposed by Conzelmann and Sandhoff.
authors.affiliation: Developmental and Metabolic Neurology Branch, NINDS, NIH, Bethesda, Maryland USA.
authors.forename: U H
authors.initials: UH
authors.lastname: Schueler
authors.forename: T
authors.initials: T
authors.lastname: Kolter
authors.forename: C R
authors.initials: CR
authors.lastname: Kaneski
authors.forename: G C
authors.initials: GC
authors.lastname: Zirzow
authors.forename: K
authors.initials: K
authors.lastname: Sandhoff
authors.forename: R O
authors.initials: RO
authors.lastname: Brady
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000378:metabolism; D005776:Gaucher Disease / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism*; D005962:Glucosylceramidase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D005963:Glucosylceramides / Q000378:metabolism; D006579:Heterozygote; D007294:Inositol / Q000031:analogs & derivatives* / Q000494:pharmacology; D050356:Lipid Metabolism; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D008854:Microscopy, Electron
chemical_list: D004791:Enzyme Inhibitors; D005963:Glucosylceramides; D007294:Inositol; D005962:Glucosylceramidase; C011423:conduritol epoxide