Rivista
/
Neuroscienze
/
Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics
JoVE Journal
Neuroscienze
È necessario avere un abbonamento a JoVE per visualizzare questo.  Accedi o inizia la tua prova gratuita.
JoVE Journal Neuroscienze
Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics

Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics

DOI:
10.3791/64132-v

•

11:40 min

•

June 23, 2022

•

, ,
1Department of Chemistry,The George Washington University

Capitoli

  • 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

Traduzione automatica

Traduzione automatica

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

Article

Embed

AGGIUNGI ALLA PLAYLIST

Usage Statistics

Video correlati

check_url/it/64132?article_type=v

DiOLISTIC Labeling of Neurons from Rodent and Non-human Primate Brain Slices
check_url/it/64132?article_type=v

Labeling F-actin Barbed Ends with Rhodamine-actin in Permeabilized Neuronal Growth Cones
check_url/it/64132?article_type=v

Recording Large-scale Neuronal Ensembles with Silicon Probes in the Anesthetized Rat
check_url/it/64132?article_type=v

Fast Micro-iontophoresis of Glutamate and GABA: A Useful Tool to Investigate Synaptic Integration
check_url/it/64132?article_type=v

Retrograde Labeling of Retinal Ganglion Cells in Adult Zebrafish with Fluorescent Dyes
check_url/it/64132?article_type=v

Preparation of Neuronal Co-cultures with Single Cell Precision
check_url/it/64132?article_type=v

Electrophysiological and Morphological Characterization of Neuronal Microcircuits in Acute Brain Slices Using Paired Patch-Clamp Recordings
check_url/it/64132?article_type=v

Extracellular Recording of Neuronal Activity Combined with Microiontophoretic Application of Neuroactive Substances in Awake Mice
check_url/it/64132?article_type=v

Quantification of Endosome and Lysosome Motilities in Cultured Neurons Using Fluorescent Probes
check_url/it/64132?article_type=v

Chronic Transcranial Electrical Stimulation and Intracortical Recording in Rats

Read Article