2. Academic Validation
  3. Glucosylated cholesterol in mammalian cells and tissues: formation and degradation by multiple cellular β-glucosidases

Glucosylated cholesterol in mammalian cells and tissues: formation and degradation by multiple cellular β-glucosidases

  • J Lipid Res. 2016 Mar;57(3):451-63. doi: 10.1194/jlr.M064923.
André R A Marques 1,  Mina Mirzaian 2,  Hisako Akiyama 3,  Patrick Wisse 4,  Maria J Ferraz 1,  Paulo Gaspar 1,  Karen Ghauharali-van der Vlugt 1,  Rianne Meijer 2,  Pilar Giraldo 5,  Pilar Alfonso 5,  Pilar Irún 5,  Maria Dahl 6,  Stefan Karlsson 6,  Elena V Pavlova 7,  Timothy M Cox 7,  Saskia Scheij 1,  Marri Verhoek 2,  Roelof Ottenhoff 1,  Cindy P A A van Roomen 1,  Navraj S Pannu 2,  Marco van Eijk 2,  Nick Dekker 1,  Rolf G Boot 2,  Herman S Overkleeft 4,  Edward Blommaart 1,  Yoshio Hirabayashi 3,  Johannes M Aerts 8
Affiliations

Affiliations

  • 1 Department of Medical Biochemistry, Academic Medical Center, Amsterdam, The Netherlands.
  • 2 Departments of Medical Biochemistry Leiden Institute of Chemistry, Leiden, The Netherlands.
  • 3 Brain Science Institute, RIKEN, Wako, Japan.
  • 4 Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden, The Netherlands.
  • 5 Centro de Investigación Biomédica en Red de Enfermedades Raras, Unidad de Investigación Traslacional, Zaragoza, Spain.
  • 6 Department of Molecular Medicine and Gene Therapy, Lund University, Lund, Sweden.
  • 7 Addenbrooke's Hospital, Department of Medicine, University of Cambridge, Cambridge, UK.
  • 8 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.
PMID: 26724485 DOI: 10.1194/jlr.M064923
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.

Keywords

Gaucher disease; Niemann-Pick type C disease; glucocerebrosidase; glucosyl-β-D-cholesterol; glucosylceramide.

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