human full-length wild-type gcase protein (cerezyme)  (Genzyme)

Journal: Scientific Reports

Article Title: A novel allosteric GCase modulator prevents Tau accumulation in GBA1 WT and GBA1 L444P/L444P cellular models

doi: 10.1038/s41598-025-02346-8

Figure Lengend Snippet: ( A ) Molecular structure of the GCase pharmacological chaperone GT-02216. ( B ) SPR dose-response for GT-02216 binding to immobilized human GCase protein monitored at acidic (pH 5.0) and neutral (pH 7.4) conditions. ( C ) SPR binding properties determined at the indicated pH values.

Article Snippet: Human full-length wild-type GCase protein (Cerezyme, Genzyme, Naarden, NL) was immobilized on the SPR CM5 sensor (GE Healthcare, #29149603,) by standard amino coupling using relatively high protein concentration of 100 μg/mL.

Techniques: Binding Assay

Journal: Scientific Reports

Article Title: A novel allosteric GCase modulator prevents Tau accumulation in GBA1 WT and GBA1 L444P/L444P cellular models

doi: 10.1038/s41598-025-02346-8

Figure Lengend Snippet: ( A ) Basal GCase activity in wild-type and mutant GBA1 fibroblasts (mean ± SD normalized on GBA1 WT(XY) , n = 6–48). Ordinary 1way ANOVA ( p < .0001) and Dunnet’s multiple tests against GBA1 WT(XY) , **** p < .0001. ( B ) Effect of 4-days treatment with 12.5 µmol/L GT-02216 on GCase activity on fibroblast lines (mean ± SD normalized on the respective basal activity, n = 5–42). Ordinary 1way ANOVA ( p < .0001) and Šidák’s multiple tests against the respective basal activity, **** p < .0001. (C) GT-02216 dose-response on GBA1 L444P/L444P(I) α fibroblasts (mean ± sem normalized on vehicle, n = 14) or (D) on GBA1 WT (XY) fibroblasts (mean ± sem normalized on vehicle, n = 8) treated for 4 days. Non-linear fit with four parameters log(agonist); EC 50 = 2.4 µmol/L on GBA1 L444P/L444P(I) α (1.3–10.5 µmol/L 95% confidentiality interval, top 1.8–2.4 µmol/L, R 2 = 0.71) and 1.5 µmol/L on GBA1 WT(XY) (1.3–1.8 µmol/L 95% confidentiality interval, top 1.9-2.0 µmol/L, R 2 = 0.95).

Article Snippet: Human full-length wild-type GCase protein (Cerezyme, Genzyme, Naarden, NL) was immobilized on the SPR CM5 sensor (GE Healthcare, #29149603,) by standard amino coupling using relatively high protein concentration of 100 μg/mL.

Techniques: Activity Assay, Mutagenesis

Journal: Scientific Reports

Article Title: A novel allosteric GCase modulator prevents Tau accumulation in GBA1 WT and GBA1 L444P/L444P cellular models

doi: 10.1038/s41598-025-02346-8

Figure Lengend Snippet: ( A ) GCase activity in doxycycline-inducible Tau-mCherry human fibroblasts with the genotype GBA WT(XX) or GBA1 L444P/L444P(I) α (mean ± SD normalized on GBA WT(XX) , n = 7). Mann-Whitney test, *** p 0.0006. ( B ) GT-02216 dose-response on Tau- GBA1 L444P/L444P(I) α fibroblasts (mean ± sem normalized on vehicle, n = 8) treated for 4 days. Non-linear fit with four parameters log(agonist); EC 50 = 1.1 µmol/L (0.7–1.6 µmol/L 95% confidentiality interval, top 1.5–1.6 µmol/L, R 2 = 0.79). ( C ) Basal and GT-02166-rescued GCase activity is not affected by the induction of Tau-mCherry expression with doxycycline in Tau-GBA1 WT(XX) fibroblasts (mean ± SD normalized on basal ctrl, n = 3). 2way ANOVA ( p < .0001 for treatment, ns for Tau expression) and Šidák’s multiple tests against the respective ctrl, **** p < .0001. ( D ) GT-02216 dose-response on Tau-GBA1 WT(XX) fibroblasts (mean ± sem normalized on vehicle, n = 8) treated for 4 days. Non-linear fit with four parameters log(agonist); EC 50 = 1.0 µmol/L (0.7–2.3 µmol/L 95% confidentiality interval, top 1.4–1.6 µmol/L, R 2 = 0.78).

Article Snippet: Human full-length wild-type GCase protein (Cerezyme, Genzyme, Naarden, NL) was immobilized on the SPR CM5 sensor (GE Healthcare, #29149603,) by standard amino coupling using relatively high protein concentration of 100 μg/mL.

Techniques: Activity Assay, MANN-WHITNEY, Expressing

Journal: Scientific Reports

Article Title: A novel allosteric GCase modulator prevents Tau accumulation in GBA1 WT and GBA1 L444P/L444P cellular models

doi: 10.1038/s41598-025-02346-8

Figure Lengend Snippet: ( A ) Quantification of Tau Puncta (TP) in Tau-GBA1 WT(XX) and Tau-GBA1 L444P/L444P(I) α fibroblasts at basal conditions (mean ± sem normalized on Tau-GBA WT(XX) , n = 55–60). Mann-Whitney test, *** p 0.0004. ( B ) Quantification of Tau puncta in Tau-GBA1 L444P/L444P(I) α fibroblasts treated overnight in the absence (ctrl) or presence (seeds) of Alzheimer’s brain-derived Tau seeds (mean ± sem normalized on ctrl, n = 45). Mann-Whitney test, **** p 4 × 10 − 11 . A representative image of ctrl or Tau seeds conditions are shown (Tau in magenta, nuclei stained with DAPI in blue). ( C ) GCase activity in Tau-GBA1 L444P/L444P(I) α fibroblasts treated overnight in the absence or presence of Tau seeds (mean ± sem normalized on ctrl, n = 3). Mann-Whitney test, not significant. ( D ) as in B. for Tau-GBA1 WT(XX) fibroblasts ( n = 45). Mann-Whitney test, **** p 0.00001. ( E ) as in C. for Tau-GBA1 WT(XX) fibroblasts ( n = 6–9). Mann-Whitney test, not significant. ( F ) Dose-dependent reduction of Tau Puncta in Tau- GBA1 L444P/L444P(I) α fibroblasts or ( G ) in GBA1 WT(XX) fibroblasts treated for 4 days with the indicated amount of GT-02216 (mean ± sem normalized on vehicle, no seed ctrl, n = 15–45). 2way ANOVA ( p < .0001 for GT-02216 concentration and seeds) and Šídák’s multiple tests against vehicle, **** p < .0001.

Article Snippet: Human full-length wild-type GCase protein (Cerezyme, Genzyme, Naarden, NL) was immobilized on the SPR CM5 sensor (GE Healthcare, #29149603,) by standard amino coupling using relatively high protein concentration of 100 μg/mL.

Techniques: MANN-WHITNEY, Derivative Assay, Staining, Activity Assay, Concentration Assay