TY - JOUR
T1 - Glucosylated cholesterol in mammalian cells and tissues: formation and degradation by multiple cellular β-glucosidases
AU - Marques, André R. A.
AU - Mirzaian, Mina
AU - Akiyama, Hisako
AU - Wisse, Patrick
AU - Ferraz, Maria J.
AU - Gaspar, Paulo
AU - Ghauharali-van der Vlugt, Karen
AU - Meijer, Rianne
AU - Giraldo, Pilar
AU - Alfonso, Pilar
AU - Irún, Pilar
AU - Dahl, Maria
AU - Karlsson, Stefan
AU - Pavlova, Elena V.
AU - Cox, Timothy M.
AU - Scheij, Saskia
AU - Verhoek, Marri
AU - Ottenhoff, Roelof
AU - van Roomen, Cindy P. A. A.
AU - Pannu, Navraj S.
AU - van Eijk, Marco
AU - Dekker, Nick
AU - Boot, Rolf G.
AU - Overkleeft, Herman S.
AU - Blommaart, Edward
AU - Hirabayashi, Yoshio
AU - Aerts, Johannes M.
PY - 2016
Y1 - 2016
N2 - 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
AB - 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
U2 - https://doi.org/10.1194/jlr.M064923
DO - https://doi.org/10.1194/jlr.M064923
M3 - Article
C2 - 26724485
SN - 0022-2275
VL - 57
SP - 451
EP - 463
JO - Journal of Lipid Research
JF - Journal of Lipid Research
IS - 3
ER -