Journal
/
Biochimie
/
Isolation of Physiologically Active Thylakoids and Their Use in Energy-Dependent Protein Transport Assays
Journal JoVE
Biochimie
Un abonnement à JoVE est nécessaire pour voir ce contenu.  Connectez-vous ou commencez votre essai gratuit.
Journal JoVE Biochimie
Isolation of Physiologically Active Thylakoids and Their Use in Energy-Dependent Protein Transport Assays

Isolation of Physiologically Active Thylakoids and Their Use in Energy-Dependent Protein Transport Assays

DOI:
10.3791/58393-v

•

12:25 min

•

September 28, 2018

•

, , ,
1Department of Plant Biology,University of California – Davis

Chapitres

  • 00:04Titre
  • 00:30Chloroplast Isolation and Quantification
  • 03:30Isolation of Thylakoids
  • 04:42Stromal Extract (SE) Recovery
  • 06:33Transport through the cpTat Pathway
  • 07:44Transport through the cpSec1 and cpSRP Pathways
  • 08:59Thermolysin Treatment
  • 10:16Results: Transport of Proteins Through cpTat and cpSec1 and Integration Through cpSRP Pathways
  • 11:18Conclusion

Summary

Traduction automatique

Traduction automatique

We present protocols herein for high-yield isolation of physiologically active thylakoids and protein transport assays for the chloroplast twin arginine translocation (cpTat), secretory (cpSec1), and signal recognition particle (cpSRP) pathways.

Tags

ThylakoidsProtein TransportChloroplast MembranesProtein TargetingPercoll GradientChloroplast IsolationPea ShootsHomogenizationCentrifugationChlorophyll Concentration

Article

Embed

AJOUTER À LA PLAYLIST

Usage Statistics

Vidéos Connexes

An Efficient Method for the Isolation of Highly Purified RNA from Seeds for Use in Quantitative Transcriptome Analysis

Synthesis of 1,2-Azaborines and the Preparation of Their Protein Complexes with T4 Lysozyme Mutants

Analyzing Supercomplexes of the Mitochondrial Electron Transport Chain with Native Electrophoresis, In-gel Assays, and Electroelution

Biochemical and Structural Characterization of the Carbohydrate Transport Substrate-binding-protein SP0092

Isolation and Respiratory Measurements of Mitochondria from Arabidopsis thaliana

Split-BioID — Proteomic Analysis of Context-specific Protein Complexes in Their Native Cellular Environment

Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation

Analysis of Protein Folding, Transport, and Degradation in Living Cells by Radioactive Pulse Chase

In Vitro Ubiquitination and Deubiquitination Assays of Nucleosomal Histones

Application of Biolayer Interferometry (BLI) for Studying Protein-Protein Interactions in Transcription

Read Article