Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

6.9K
Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
6.9K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

[Influence of sampling satisfaction using endometrial sampling device and related factors for pathology diagnostic accordance rate].

Zhonghua fu chan ke za zhi·2014
Same author

Effect of the number of positive lymph nodes and lymph node ratio on prognosis of patients after resection of pancreatic adenocarcinoma.

Hepatobiliary & pancreatic diseases international : HBPD INT·2014
Same author

Mechanisms of human erythrocytic bioactivation of nitrite.

The Journal of biological chemistry·2014
Same author

Simulation research on the natural degradation process of PBDEs in soil polluted by e-waste under increased concentrations of atmospheric O(3).

Chemosphere·2014
Same author

Two-dimensional colloidal crystal assisted formation of conductive porous gold films with flexible structural controllability.

Journal of colloid and interface science·2014
Same author

REDD1 attenuates cardiac hypertrophy via enhancing autophagy.

Biochemical and biophysical research communications·2014

Related Experiment Video

Updated: Sep 16, 2025

AirID-Based Proximity Labeling for Protein-Protein Interaction in Plants
08:36

AirID-Based Proximity Labeling for Protein-Protein Interaction in Plants

Published on: September 16, 2022

1.8K

Coupling Affinity Purification with Proximity Labeling in Arabidopsis: The APEAL Approach.

Chen Liu1, Panagiotis N Moschou2,3,4

  • 1State Key Laboratory of Biocontrol, Guangdong Key Laboratory of Plant Resources, School of Life Sciences, Sun Yat-Sen University, Guangzhou, China.

Methods in Molecular Biology (Clifton, N.J.)
|July 10, 2025
PubMed
Summary

We developed APEAL (Tandemly Coupled Affinity Purification with Proximity-dEpendent LigAtion), a new method to identify transient and stable protein interactions in plants. This approach uses TurboID for efficient protein complex capture and analysis via mass spectrometry.

Keywords:
APEALProtein pull downProximity-dependent ligationTurboID

More Related Videos

Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics
11:40

Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics

Published on: June 23, 2022

2.6K
2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes
08:23

2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes

Published on: August 6, 2018

11.5K

Related Experiment Videos

Last Updated: Sep 16, 2025

AirID-Based Proximity Labeling for Protein-Protein Interaction in Plants
08:36

AirID-Based Proximity Labeling for Protein-Protein Interaction in Plants

Published on: September 16, 2022

1.8K
Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics
11:40

Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics

Published on: June 23, 2022

2.6K
2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes
08:23

2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes

Published on: August 6, 2018

11.5K

Area of Science:

  • Plant molecular biology
  • Proteomics
  • Biochemistry

Background:

  • Identifying transient protein-protein interactions is crucial for understanding cellular processes.
  • Existing methods often struggle to capture weak or short-lived protein assemblies.
  • Plant systems present unique challenges for studying protein interactions due to their cellular environment.

Purpose of the Study:

  • To introduce and detail the APEAL method for capturing both transient and stable protein assemblies in plants.
  • To provide a comprehensive protocol for implementing APEAL, from experimental design to data analysis.
  • To enable researchers to study dynamic protein interactions in various plant species.

Main Methods:

  • APEAL combines affinity purification for strong interactions with proximity-dependent ligation for transient interactions.
  • Utilizes TurboID, a highly efficient biotin ligase variant, operating at plant-friendly temperatures.
  • Analysis of captured protein complexes is performed using immunoblotting or mass spectrometry.

Main Results:

  • APEAL successfully captures a spectrum of protein interactions, from stable to transient.
  • The method is applicable to diverse plant species, including Arabidopsis.
  • Detailed protocols are provided for construct design, optimization, and data analysis.

Conclusions:

  • APEAL offers a robust and versatile approach for mapping protein interaction networks in plants.
  • The method enhances the ability to study dynamic protein complexes, advancing plant science.
  • This technique facilitates deeper insights into plant cellular mechanisms and responses.