Collection of protocols of Team Deleidi used in the publication: “”LRRK2 kinase activity regulates GCase level and enzymatic activity differently depending on cell type in Parkinson’s disease””

By Federico Bertoli, MD and Michela Deleidi, MD, PhD
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Output Details

Description

Collection of protocols of Team Deleidi used in the publication: ""LRRK2 kinase activity regulates GCase level and enzymatic activity differently depending on cell type in Parkinson’s disease""

Tags
  • Activity Assay
  • GBA (Glucocerebrosidase)
  • GCase activity
Team
Team Schapira
Program
Collaborative Research Network
Theme
Neuro-Immune Interactions

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LRRK2 kinase activity regulates GCase level and enzymatic activity differently depending on cell type in Parkinson’s disease

Leucine-rich repeat kinase 2 (LRRK2) is a kinase involved in different cellular functions, including autophagy, endolysosomal pathways, and immune function. Mutations in LRRK2 cause autosomal-dominant forms of Parkinson’s disease (PD).…

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Lysosomal GCase (glucocerebrosidase) activity assay

Method to measure lysosomal GBA1 enzyme activity using 4-MUG substrate at low pH to minimize non-lysosomal GCase interference. GBA1 activity higher in purified lysosomes than whole cell extract, confirmed by CBE inhibitor abolishing 4-MUG hydrolysis.

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Ex Vivo Mouse Brain Slice Whole-Cell Patch-Clamp Electrophysiology

This protocol outlines steps for performing ex vivo slice whole-cell patch-clamp electrophysiology experiments, including voltage clamp and current clamp recordings as well as optogenetic stimulation in the Ding lab.

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