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

Mapping Dysfunctional Protein-Protein Interactions in Disease09:39

Mapping Dysfunctional Protein-Protein Interactions in Disease

607
Here, we present a protocol to enable the capture and identification of disease-specific protein-protein interactions from native cells and tissues using chemical probes and mass spectrometry. The resulting interaction datasets are analyzed through a dedicated web-based platform to reveal dynamic network dysfunctions and pathway alterations linked to...
607
Split-BioID — Proteomic Analysis of Context-specific Protein Complexes in Their Native Cellular Environment09:02

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

20.4K
We provide a step-by-step protocol for split-BioID, a protein fragments-complementation assay based on the proximity-labeling technique BioID. Activated on the interaction of two given proteins, it allows the proteomics analysis of context-dependent protein complexes in their native cellular environment. The method is simple, cost-effective and only requires standard laboratory...
20.4K
Identifying Protein-protein Interaction Sites Using Peptide Arrays07:44

Identifying Protein-protein Interaction Sites Using Peptide Arrays

18.5K
Peptide array screening is a high throughput assay for identifying protein-protein interaction sites. This allows mapping multiple interactions of a target protein and can serve as a method for identifying sites for inhibitors that target a protein. Here we describe a protocol for screening and analyzing peptide...
18.5K
Study of Protein-protein Interactions in Autophagy Research14:08

Study of Protein-protein Interactions in Autophagy Research

9.5K
Presented here are two antibody-based protein-protein interaction research techniques: immunofluorescence and immunoprecipitation. These techniques are suitable for studying physical interactions between proteins for the discovery of novel components of cellular signaling pathways and for understanding protein dynamics.
9.5K
Imaging Protein-protein Interactions in vivo11:15

Imaging Protein-protein Interactions in vivo

21.8K
This protocol describes how to image protein-protein interactions using a FRET-based proximity...
21.8K
Microtubule Associated Proteins (MAPs)01:42

Microtubule Associated Proteins (MAPs)

5.8K
Microtubule function and architecture are regulated by an array of specialized proteins called microtubule-associated proteins or MAPs. These proteins are widespread across different organisms and have conserved protein motifs, like the multi-TOG domain for tubulin binding found in the CLASP family of MAPs. Some MAPs are lineage-specific based on their conserved domains. Their functions depend upon the cytoskeletal architecture and cell type they are located within. In-plant cells, a specific...
5.8K

You might also read

Related Articles

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

Sort by
Same author

Modeling Hereditary Angioedema With Personalized EPSC-Derived Hepatocytes: A CRISPR-Validated Platform for Mutation-Specific Mechanisms and Therapeutic Innovation.

Allergy·2026
Same author

Telerehabilitation versus face-to-face pulmonary rehabilitation in COPD: A systematic review and meta-analysis of comparative outcomes and delivery characteristics.

Journal of telemedicine and telecare·2026
Same author

Continental-wide prioritization of protected areas to address global and habitat-specific goals for avian biodiversity conservation.

Conservation biology : the journal of the Society for Conservation Biology·2026
Same author

Electric Scooters: A Modern Method of Trauma.

Missouri medicine·2026
Same author

Surgery for interplanetary space missions.

The British journal of surgery·2026
Same author

Sulfonyl Anthranilic Acid Analogues Display Pan-Serotype Anti-Dengue Activity by Downregulating the Expression of Ribosomal Proteins Encoded by 5'-Terminal Oligopyrimidine Motif-Containing mRNA.

Journal of medicinal chemistry·2026
Same journal

Measuring Mitochondrial Respiration in Previously Frozen Biological Samples.

Current protocols in cell biology·2020
Same journal

Proximity Ligation Assay for Detecting Protein-Protein Interactions and Protein Modifications in Cells and Tissues in Situ.

Current protocols in cell biology·2020
Same journal

Methods for Investigating Corneal Cell Interactions and Extracellular Vesicles In Vitro.

Current protocols in cell biology·2020
Same journal

Multiplexed Proximity Biotinylation Coupled to Mass Spectrometry for Defining Integrin Adhesion Complexes.

Current protocols in cell biology·2020
Same journal

Preparation of Extracellular Matrix Paper and Construction of Multi-Layered 3D Tissue Model.

Current protocols in cell biology·2020
Same journal

Metabolic Analysis at the Nanoscale with Multi-Isotope Imaging Mass Spectrometry (MIMS).

Current protocols in cell biology·2020
See all related articles

Related Experiment Video

Updated: Jan 20, 2026

Mapping Dysfunctional Protein-Protein Interactions in Disease
09:39

Mapping Dysfunctional Protein-Protein Interactions in Disease

Published on: October 24, 2025

607

Protein-Protein Interaction Mapping by 2C-BioID.

Alexandre Chojnowski1, Hendrikje Werner1, Matthew Cook1

  • 1Developmental and Regenerative Biology, Institute of Medical Biology, Agency for Science, Technology and Research (A*STAR), Singapore, Singapore.

Current Protocols in Cell Biology
|September 5, 2019
PubMed
Summary
This summary is machine-generated.

Researchers developed 2C-BioID, a new method to study protein-protein interactions (PPIs). This technique improves the specificity of BioID assays by controlling when the biotin ligase and protein of interest interact, reducing false positives.

Keywords:
2C-BioIDAP21967BioIDFKBPFRBPPI

More Related Videos

Split-BioID — Proteomic Analysis of Context-specific Protein Complexes in Their Native Cellular Environment
09:02

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

Published on: April 20, 2018

20.4K
Identifying Protein-protein Interaction Sites Using Peptide Arrays
07:44

Identifying Protein-protein Interaction Sites Using Peptide Arrays

Published on: November 18, 2014

18.5K

Related Experiment Videos

Last Updated: Jan 20, 2026

Mapping Dysfunctional Protein-Protein Interactions in Disease
09:39

Mapping Dysfunctional Protein-Protein Interactions in Disease

Published on: October 24, 2025

607
Split-BioID — Proteomic Analysis of Context-specific Protein Complexes in Their Native Cellular Environment
09:02

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

Published on: April 20, 2018

20.4K
Identifying Protein-protein Interaction Sites Using Peptide Arrays
07:44

Identifying Protein-protein Interaction Sites Using Peptide Arrays

Published on: November 18, 2014

18.5K

Area of Science:

  • Molecular Biology
  • Cellular Biology
  • Biochemistry

Background:

  • Protein-protein interactions (PPIs) are crucial for cellular functions and signal transduction.
  • Existing methods like BioID assay interrogate PPIs in vivo but have limitations.
  • Improving the specificity and reducing false positives in PPI studies is essential.

Purpose of the Study:

  • To develop an improved proximity-dependent biotinylation assay for studying PPIs.
  • To enhance the specificity of BioID by controlling enzyme-target proximity.
  • To reduce false-positive interactome data.

Main Methods:

  • Developed 2C-BioID, a modified BioID assay utilizing the FKBP-FRB dimerization system.
  • Induced proximity between biotin ligase and the protein of interest using rapamycin analogue AP21967.
  • Compared 2C-BioID with standard BioID for interactome analysis.

Main Results:

  • 2C-BioID ensures biotin ligase association only upon addition of AP21967.
  • The method effectively alleviates targeting issues associated with standard BioID.
  • 2C-BioID demonstrates improved specificity in identifying protein interactomes, reducing false positives.

Conclusions:

  • 2C-BioID offers a refined approach for studying PPIs within a cellular context.
  • This method enhances the accuracy and reliability of interactome mapping.
  • 2C-BioID represents a valuable advancement for researchers investigating cellular signaling pathways.