Effect of Expression of Human Glucosylceramidase 2 Isoforms on Lipid Profiles in COS-7 Cells
Peeranat Jatooratthawichot, Chutima Talabnin, Lukana Ngiwsara, Yepy Hardi Rustam, Jisnuson Svasti, Gavin E Reid, James R Ketudat Cairns
Metabolites | MDPI AG | Published : 2020
Glucosylceramide (GlcCer) is a major membrane lipid and the precursor of gangliosides. GlcCer is mainly degraded by two enzymes, lysosomal acid β-glucosidase (GBA) and nonlysosomal β-glucosidase (GBA2), which may have different isoforms because of alternative splicing. To understand which GBA2 isoforms are active and how they affect glycosphingolipid levels in cells, we expressed nine human GBA2 isoforms in COS-7 cells, confirmed their expression by qRT-PCR and Western blotting, and assayed their activity to hydrolyze 4-methylumbelliferyl-β-D-glucopyranoside (4MUG) in cell extracts. Human GBA2 isoform 1 showed high activity, while the other isoforms had activity similar to the background. Co..View full abstract
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Awarded by Australian Research Council
Financial support was provided by Suranaree University of Technology, including the National Research University Project of the Commission on Higher Education, and also from the Australian Research Council to GER (LE160100015). PJ was supported by a Development and a Promotion of Science and Technology Talents Project scholarship.