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| Solution Information | help | |
| Enzyme: | Lysosomal acid glucosylceramidase | |
| inhibitor: | BDBM42394 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: The purpose of this assay is to determine dose response curves for test compounds identified as active in a previous screen entitled, "Primary Cell-based high throughput screening assay for enhancers of beta-glucosidase activity," (PubChem AID 784). In this assay, activation of beta-glucosidase was measured using fibroblasts cells derived from a Gaucher disease patient with the homozygous L444P beta-glucosidase mutation. These cells have significantly reduced lysosomal glucosidase activity as a result of misfolding and degradation of the mutant protein, which reduces the amount of enzyme in the lysosome. Cells are treated with test compounds for 3 days to allow the enzyme to accumulate in the lysosome, followed by treatment with the fluorescent beta-glucosidase substrate 4-Methylumbelliferyl-B-D-glucopyranoside (MUD). As designed, a compound that acts as a beta-glucosidase activator will increase cleavage of MUD, and as a result increase well fluorescence. Compou | |
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