Activation and Purification of ß‐Glucocerebrosidase by Exploiting its Transporter LIMP‐2 – Implications for Novel Treatment Strategies in Gaucher's and Parkinson's Disease

Advanced Science, Год журнала: 2024, Номер 11(25)

Опубликована: Апрель 26, 2024

Abstract Genetic variants of GBA1 can cause the lysosomal storage disorder Gaucher disease and are among highest genetic risk factors for Parkinson's (PD). encodes enzyme beta‐glucocerebrosidase (GCase), which orchestrates degradation glucosylceramide (GluCer) in lysosome. Recent studies have shown that GluCer accelerates α ‐synuclein aggregation, exposing GCase deficiency as a major factor PD pathology promising target treatment. This study investigates interaction three disease‐associated (p.E326K, p.N370S, p.L444P) with their transporter, integral membrane protein 2 (LIMP‐2). Overexpression LIMP‐2 HEK 293T cells boosts abundance wt, E326K, N370S increases/rescues enzymatic activity wt E326K variant. Using novel purification approach, co‐purification untagged complex His‐tagged from cell supernatant 293F is achieved, confirming functional binding trafficking these variants. Furthermore, single helix ectodomain exploited to design lysosome‐targeted peptide enhances patient‐derived control fibroblasts. These findings reveal an allosteric activator GCase, suggesting possible therapeutic potential targeting this interaction.

Язык: Английский

Cryo-TEM structure of β-glucocerebrosidase in complex with its transporter LIMP-2

Research Square (Research Square), Год журнала: 2024, Номер unknown

Опубликована: Апрель 29, 2024

Nearly all lysosomal enzymes reach the lysosome via mannose-6-phosphate receptor pathway. One of few known exceptions is enzyme β-glucocerebrosidase (GCase) that requires integral membrane protein type-2 (LIMP-2) as a proprietary transporter. Genetic variations in GCase encoding gene GBA1 are cause Gaucher’s disease (GD) and highest genetic risk factor for developing Parkinson’s (PD). Activators targeting have emerged promising therapeutic approach treatment both GD PD, with pre-clinical clinical trials ongoing. In this study, we resolved complex LIMP-2 to 3.6 Å using cryo-electron microscopy aid an engineered shuttle two GCase-targeted pro-macrobodies. We identified helix 5 7 interact binding pocket GCase, forming mostly hydrophobic interaction interface. Understanding interplay on structural level crucial identifying potential activation sites conceptualizing novel approaches GCase. Here, unveil structure mannose-6-phosphate-independent transporting provide fundamental knowledge translational research overcome PD.

Язык: Английский

Engineering synthetic and recombinant human lysosomal β-glucocerebrosidase for enzyme replacement therapy for Gaucher disease

Deleted Journal, Год журнала: 2024, Номер 6(10)

Опубликована: Окт. 4, 2024

Язык: Английский