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Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics
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Neuroscience
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JoVE Journal Neuroscience
Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics

Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics

DOI:
10.3791/64132-v

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11:40 min

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June 23, 2022

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, ,
1Department of Chemistry,The George Washington University

Chapters

  • 00:05Introduction
  • 00:59Proximity Labeling in Human iPSC-Derived Neurons
  • 03:17Enriching Biotinylated Proteins and On-Beads Digestion
  • 06:39LC-MS Analysis
  • 07:15Proteomics Data Analysis
  • 08:36Results: Lysosome Proximity Labeling Proteomics Study Conducted in Human iPSC-Derived Neurons
  • 10:43Conclusion

Summary

Automatic Translation

Automatic Translation

A neuronal lysosome proximity labeling proteomics protocol is described here to characterize the dynamic lysosomal microenvironment in human induced pluripotent stem cell-derived neurons. Lysosomal membrane proteins and proteins that interact with lysosomes (stably or transiently) can be accurately quantified in this method with excellent intracellular spatial resolution in live human neurons.

Tags

Keywords: Proximity Labeling ProteomicsLysosomal MicroenvironmentIPSC-derived NeuronsLysosomal DysfunctionBrain DiseasesBiotin PhenolHydrogen PeroxideAcetone PrecipitationProtein PelletDCA Assay

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