Glucocerebrosidase is imported into mitochondria and preserves complex I integrity and energy metabolism – ASAP protocol collection

By Michela Deleidi, MD, PhD, Pascale Baden, MSc, Maria Jose Perez Jimenez, PhD, Hariam Raji, MSc and Federico Bertoli, MD
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Output Details

Description
Collection of protocols of Deleidi Lab used in the publication: "Glucocerebrosidase is imported into mitochondria and preserves complex I integrity and energy metabolism"
Identifier (DOI)
10.17504/protocols.io.x54v9d44mg3e/v1
Tags
  • Activity Assay
  • GCase activity
  • Immunofluorescence
  • Immunoprecipitation
  • Lentivirus
  • Western Blot
Team
Team Schapira
Program
Collaborative Research Network
Theme
Neuro-Immune Interactions

Meet the Authors

  • Michela Deleidi, MD, PhD

    Co-PI (Core Leadership): Team Schapira, 

    Institut Imagine

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    Pascale Baden, MSc

    Key Personnel: Team Schapira, 

    Deutsches Zentrum für Neurodegenerative Erkrankungen (DZNE)

  • User avatar fallback logo

    Maria Jose Perez Jimenez, PhD

    Key Personnel: Team Schapira, 

    German Center for Neurodegenerative Diseases

  • Hariam Raji, MSc

    Key Personnel: Team Schapira, 

    Institut des Maladies Génétiques Imagine

  • Federico Bertoli, MD

    Key Personnel: Team Schapira, 

    German Center for Neurodegenerative Diseases

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Glucocerebrosidase is imported into mitochondria and preserves complex I integrity and energy metabolism

Mutations in GBA1, the gene encoding the lysosomal enzyme β-glucocerebrosidase (GCase), which cause Gaucher’s disease, are the most frequent genetic risk factor for Parkinson’s disease (PD). Here, we employ global proteomic and single-cell…

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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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