title: Selective chaperone effect of aminocyclitol derivatives on G202R and other mutant glucocerebrosidases causing Gaucher disease.
abstract: Gaucher disease is an autosomal recessive lysosomal disorder characterized by the accumulation of glucosylceramide as a result of a deficiency of the enzyme glucocerebrosidase. Several competitive glucocerebrosidase inhibitors are able to act as pharmacological chaperones for an efficient rescue of the mutated, misfolded forms of the enzyme. Along this line, we report in this work on the ability of several aminocyclitols to increase the residual glucocerebrosidase activity in patient fibroblasts with different genotypes. Some of the compounds were slightly active on fibroblasts bearing some mutations, including the highly prevalent N370S mutation. All compounds were highly active as enzyme activity enhancers on fibroblasts from Gaucher disease patients containing the G202R mutation. Moreover, using the novel tagged sphingolipid ω-azidosphingosine, a reduction in the tagged glucosylceramide accumulation was also observed for selected aminocyclitols. Attempts to explain the activity impairment observed in glucocerebrosidase bearing the G202R mutation by comparative molecular dynamic studies on wild type and the G202R mutated proteins (free and isofagomine-bound, in both cases) were unsuccessful. Under the simulation conditions used, no clear effect of the G202R mutation neither over the global structure of the protein nor on the loops that constitute the glucocerebrosidase active site was observed. Since the G202R residue is located on the protein surface, altered protein-membrane or protein-protein interactions could account for the observed differences. In conclusion, we have tested novel compounds that have shown some chaperone effect on particular glucocerebrosidase mutant enzymes, supporting the enhancement therapy as an alternative approach for Gaucher disease.
authors.affiliation: Departament de Genètica, Universitat de Barcelona (UB); IBUB; CIBER de Enfermedades Raras (CIBERER), Av. Diagonal 645, E-08028, Barcelona, Spain.
authors.forename: Jenny
authors.initials: J
authors.lastname: Serra-Vinardell
authors.affiliation: Research Unit on BioActive Molecules (RUBAM), Departament de Química Biomèdica, Institut de Química Avançada de Catalunya (IQAC-CSIC), Jordi Girona 18, 08034 Barcelona, Spain; Facultat de Farmàcia, Unitat de Química Farmacèutica (Unitat Associada al CSIC), Universitat de Barcelona (UB), Avda. Joan XXIII, s/n, 08028 Barcelona, Spain.
authors.forename: Lucía
authors.initials: L
authors.lastname: Díaz
authors.affiliation: Department of Cell and Molecular Biology, Uppsala University, Biomedical Center, Box 596, SE-751 24, Uppsala, Sweden.
authors.forename: Hugo
authors.initials: H
authors.lastname: Gutiérrez-de Terán
authors.affiliation: Departament de Genètica, Universitat de Barcelona (UB); IBUB; CIBER de Enfermedades Raras (CIBERER), Av. Diagonal 645, E-08028, Barcelona, Spain.
authors.forename: Gessamí
authors.initials: G
authors.lastname: Sánchez-Ollé
authors.affiliation: Departament de Química Biològica i Modeltzació Molecular, Institut de Química Avançada de Catalunya (IQAC-CSIC), Jordi Girona 18, 08034 Barcelona, Spain.
authors.forename: Jordi
authors.initials: J
authors.lastname: Bujons
authors.affiliation: Department of Enzymology and Cellular Function, Institute of Child Health, Athens 11527, Greece.
authors.forename: Helen
authors.initials: H
authors.lastname: Michelakakis
authors.affiliation: Department of Enzymology and Cellular Function, Institute of Child Health, Athens 11527, Greece.
authors.forename: Irene
authors.initials: I
authors.lastname: Mavridou
authors.affiliation: Department of Medicinal Biochemistry, Academic Medical Center. Meibergdreef 15, 1105 AZ, Amsterdam, The Netherlands.
authors.forename: Johannes M F G
authors.initials: JM
authors.lastname: Aerts
authors.affiliation: Research Unit on BioActive Molecules (RUBAM), Departament de Química Biomèdica, Institut de Química Avançada de Catalunya (IQAC-CSIC), Jordi Girona 18, 08034 Barcelona, Spain; Facultat de Farmàcia, Unitat de Química Farmacèutica (Unitat Associada al CSIC), Universitat de Barcelona (UB), Avda. Joan XXIII, s/n, 08028 Barcelona, Spain.
authors.forename: Antonio
authors.initials: A
authors.lastname: Delgado
authors.affiliation: Departament de Genètica, Universitat de Barcelona (UB); IBUB; CIBER de Enfermedades Raras (CIBERER), Av. Diagonal 645, E-08028, Barcelona, Spain.
authors.forename: Daniel
authors.initials: D
authors.lastname: Grinberg
authors.affiliation: Departament de Genètica, Universitat de Barcelona (UB); IBUB; CIBER de Enfermedades Raras (CIBERER), Av. Diagonal 645, E-08028, Barcelona, Spain.
authors.forename: Lluïsa
authors.initials: L
authors.lastname: Vilageliu
authors.affiliation: Research Unit on BioActive Molecules (RUBAM), Departament de Química Biomèdica, Institut de Química Avançada de Catalunya (IQAC-CSIC), Jordi Girona 18, 08034 Barcelona, Spain. Electronic address: fina.casas@iqac.csic.es.
authors.forename: Josefina
authors.initials: J
authors.lastname: Casas
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D002478:Cells, Cultured; D018360:Crystallography, X-Ray; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D005455:Fluorescent Antibody Technique; D005776:Gaucher Disease / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics; D005962:Glucosylceramidase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D050112:Imino Pyranoses / Q000494:pharmacology*; D008055:Lipids; D018613:Microscopy, Confocal; D018832:Molecular Chaperones / Q000494:pharmacology*; D056004:Molecular Dynamics Simulation; D009154:Mutation / Q000235:genetics*; D011487:Protein Conformation; D017510:Protein Folding; D012867:Skin / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013107:Sphingolipids
chemical_list: D004791:Enzyme Inhibitors; D050112:Imino Pyranoses; D008055:Lipids; D018832:Molecular Chaperones; D013107:Sphingolipids; C098432:isofagomine; D005962:Glucosylceramidase
keywords: Fibroblast; G202R mutation; Gaucher disease; Glucosylceramide; Pharmacological chaperone