title: Clinical phenotype of Gaucher disease in relation to properties of mutant glucocerebrosidase in cultured fibroblasts.
abstract: We have investigated several parameters of glucocerebrosidase in cultured skin fibroblasts from patients with various clinical phenotypes of Gaucher disease. In this study no strict correlation was found between the clinical manifestations of Gaucher disease and the parameters investigated in fibroblasts. These parameters included the specific activity of the enzyme in extracts towards natural lipid and artificial substrate in the presence of different activators; the enzymic activity per unit of glucocerebrosidase protein; the rate of synthesis of the enzyme and its stability; and the post-translational processing of the enzyme. In addition, the activity in situ of glucocerebrosidase in fibroblasts was investigated using a novel method by analysis of the catabolism of NBD-glucosylceramide in cells that were loaded with bovine serum albumin-lipid complexes. Again, no complete correlation with the clinical phenotype of patients was detectable. Glucocerebrosidase in fibroblasts from most non-neuronopathic (type 1) Gaucher disease patients differs in some aspects from enzyme in cells from patients with neurological forms (types 2 and 3). The stimulation by activator protein and phospholipid is clearly more pronounced in type 1 than in types 2 and 3; the enzymic activity per unit of glucocerebrosidase protein in type 1 is severely reduced in the presence of taurocholate and the amount of glucocerebrosidase appears (near) normal in contrast to the situation in types 2 and 3 Gaucher fibroblasts. However, this distinction was not always consistent; glucocerebrosidase in fibroblasts from some type 1 Gaucher patients, particularly some South African cases, was comparable in properties to enzyme in type 2 and 3 patients.
authors.affiliation: E.C. Slater Institute for Biochemical Research, University of Amsterdam, The Netherlands.
authors.forename: S
authors.initials: S
authors.lastname: Van Weely
authors.forename: M B
authors.initials: MB
authors.lastname: Van Leeuwen
authors.forename: I D
authors.initials: ID
authors.lastname: Jansen
authors.forename: M A
authors.initials: MA
authors.lastname: De Bruijn
authors.forename: E M
authors.initials: EM
authors.lastname: Brouwer-Kelder
authors.forename: A W
authors.initials: AW
authors.lastname: Schram
authors.forename: M C
authors.initials: MC
authors.lastname: Sa Miranda
authors.forename: J A
authors.initials: JA
authors.lastname: Barranger
authors.forename: E M
authors.initials: EM
authors.lastname: Petersen
authors.forename: J
authors.initials: J
authors.lastname: Goldblatt
mesh_terms: D002402:Cathepsin D / Q000378:metabolism; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000201:enzymology; D005776:Gaucher Disease / Q000201:enzymology* / Q000235:genetics; D005962:Glucosylceramidase / Q000235:genetics / Q000378:metabolism*; D005963:Glucosylceramides / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D009154:Mutation; D010641:Phenotype; D013696:Temperature; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
chemical_list: D005963:Glucosylceramides; D005962:Glucosylceramidase; D001619:beta-N-Acetylhexosaminidases; D002402:Cathepsin D