title: Glucosylated cholesterol in mammalian cells and tissues: formation and degradation by multiple cellular β-glucosidases.
abstract: The membrane lipid glucosylceramide (GlcCer) is continuously formed and degraded. Cells express two GlcCer-degrading β-glucosidases, glucocerebrosidase (GBA) and GBA2, located in and outside the lysosome, respectively. Here we demonstrate that through transglucosylation both GBA and GBA2 are able to catalyze in vitro the transfer of glucosyl-moieties from GlcCer to cholesterol, and vice versa. Furthermore, the natural occurrence of 1-O-cholesteryl-β-D-glucopyranoside (GlcChol) in mouse tissues and human plasma is demonstrated using LC-MS/MS and (13)C6-labeled GlcChol as internal standard. In cells, the inhibition of GBA increases GlcChol, whereas inhibition of GBA2 decreases glucosylated sterol. Similarly, in GBA2-deficient mice, GlcChol is reduced. Depletion of GlcCer by inhibition of GlcCer synthase decreases GlcChol in cells and likewise in plasma of inhibitor-treated Gaucher disease patients. In tissues of mice with Niemann-Pick type C disease, a condition characterized by intralysosomal accumulation of cholesterol, marked elevations in GlcChol occur as well. When lysosomal accumulation of cholesterol is induced in cultured cells, GlcChol is formed via lysosomal GBA. This illustrates that reversible transglucosylation reactions are highly dependent on local availability of suitable acceptors. In conclusion, mammalian tissues contain GlcChol formed by transglucosylation through β-glucosidases using GlcCer as donor. Our findings reveal a novel metabolic function for GlcCer.
authors.affiliation: Department of Medical Biochemistry, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: André R A
authors.initials: AR
authors.lastname: Marques
authors.affiliation: Departments of Medical Biochemistry Leiden Institute of Chemistry, Leiden, The Netherlands.
authors.forename: Mina
authors.initials: M
authors.lastname: Mirzaian
authors.affiliation: Brain Science Institute, RIKEN, Wako, Japan.
authors.forename: Hisako
authors.initials: H
authors.lastname: Akiyama
authors.affiliation: Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden, The Netherlands.
authors.forename: Patrick
authors.initials: P
authors.lastname: Wisse
authors.affiliation: Department of Medical Biochemistry, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Ferraz
authors.affiliation: Department of Medical Biochemistry, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Paulo
authors.initials: P
authors.lastname: Gaspar
authors.affiliation: Department of Medical Biochemistry, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Karen
authors.initials: K
authors.lastname: Ghauharali-van der Vlugt
authors.affiliation: Departments of Medical Biochemistry Leiden Institute of Chemistry, Leiden, The Netherlands.
authors.forename: Rianne
authors.initials: R
authors.lastname: Meijer
authors.affiliation: Centro de Investigación Biomédica en Red de Enfermedades Raras, Unidad de Investigación Traslacional, Zaragoza, Spain.
authors.forename: Pilar
authors.initials: P
authors.lastname: Giraldo
authors.affiliation: Centro de Investigación Biomédica en Red de Enfermedades Raras, Unidad de Investigación Traslacional, Zaragoza, Spain.
authors.forename: Pilar
authors.initials: P
authors.lastname: Alfonso
authors.affiliation: Centro de Investigación Biomédica en Red de Enfermedades Raras, Unidad de Investigación Traslacional, Zaragoza, Spain.
authors.forename: Pilar
authors.initials: P
authors.lastname: Irún
authors.affiliation: Department of Molecular Medicine and Gene Therapy, Lund University, Lund, Sweden.
authors.forename: Maria
authors.initials: M
authors.lastname: Dahl
authors.affiliation: Department of Molecular Medicine and Gene Therapy, Lund University, Lund, Sweden.
authors.forename: Stefan
authors.initials: S
authors.lastname: Karlsson
authors.affiliation: Addenbrooke's Hospital, Department of Medicine, University of Cambridge, Cambridge, UK.
authors.forename: Elena V
authors.initials: EV
authors.lastname: Pavlova
authors.affiliation: Addenbrooke's Hospital, Department of Medicine, University of Cambridge, Cambridge, UK.
authors.forename: Timothy M
authors.initials: TM
authors.lastname: Cox
authors.affiliation: Department of Medical Biochemistry, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Saskia
authors.initials: S
authors.lastname: Scheij
authors.affiliation: Departments of Medical Biochemistry Leiden Institute of Chemistry, Leiden, The Netherlands.
authors.forename: Marri
authors.initials: M
authors.lastname: Verhoek
authors.affiliation: Department of Medical Biochemistry, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Roelof
authors.initials: R
authors.lastname: Ottenhoff
authors.affiliation: Department of Medical Biochemistry, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Cindy P A A
authors.initials: CP
authors.lastname: van Roomen
authors.affiliation: Departments of Medical Biochemistry Leiden Institute of Chemistry, Leiden, The Netherlands.
authors.forename: Navraj S
authors.initials: NS
authors.lastname: Pannu
authors.affiliation: Departments of Medical Biochemistry Leiden Institute of Chemistry, Leiden, The Netherlands.
authors.forename: Marco
authors.initials: M
authors.lastname: van Eijk
authors.affiliation: Department of Medical Biochemistry, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Nick
authors.initials: N
authors.lastname: Dekker
authors.affiliation: Departments of Medical Biochemistry Leiden Institute of Chemistry, Leiden, The Netherlands.
authors.forename: Rolf G
authors.initials: RG
authors.lastname: Boot
authors.affiliation: Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden, The Netherlands.
authors.forename: Herman S
authors.initials: HS
authors.lastname: Overkleeft
authors.affiliation: Department of Medical Biochemistry, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Edward
authors.initials: E
authors.lastname: Blommaart
authors.affiliation: Brain Science Institute, RIKEN, Wako, Japan.
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Hirabayashi
authors.affiliation: Department of Medical Biochemistry, Academic Medical Center, Amsterdam, The Netherlands Departments of Medical Biochemistry Leiden Institute of Chemistry, Leiden, The Netherlands j.m.f.g.aerts@lic.leidenuniv.nl.
authors.forename: Johannes M
authors.initials: JM
authors.lastname: Aerts
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D002784:Cholesterol / Q000378:metabolism*; D005260:Female; D005776:Gaucher Disease / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D008297:Male; D051379:Mice; D009542:Niemann-Pick Diseases / Q000378:metabolism; D000067996:RAW 264.7 Cells; D001617:beta-Glucosidase / Q000378:metabolism*
chemical_list: D002784:Cholesterol; D001617:beta-Glucosidase
keywords: Gaucher disease; Niemann-Pick type C disease; glucocerebrosidase; glucosyl-β-D-cholesterol; glucosylceramide