Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Protein Networks

Protein Networks

An organism can have thousands of different proteins, and these proteins must cooperate to ensure the health of an organism. Proteins bind to other proteins and form complexes to carry out their functions. Many proteins interact with multiple other proteins creating a complex network of protein interactions.
These interactions can be represented through maps depicting protein-protein interaction networks, represented as nodes and edges. Nodes are circles that are representative of a protein,...

Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

You might also read

Related Articles

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

Sort by

Same author

Heart Failure Medication Withdrawal in Patients With Improved Cardiac Function After Atrial Fibrillation Ablation: The DEFINITION-AF Pilot Randomized Clinical Trial.

JAMA network open·2026

Same author

Aligning Chemical Kinetics with Crystallization Enables Millimeter-Scale Single Crystals of Conductive MOFs.

Journal of the American Chemical Society·2026

Same author

Anisotropic porous hydrogel fiber coating modified titanium modulates the immune microenvironment to enhance endogenous bone regeneration.

Biomaterials·2026

Same author

Development and improvement of Chinese Haemovigilance Network from 2018 to 2023.

Vox sanguinis·2026

Same author

Preprocedural hs-CRP level predicts long-term outcomes after first radiofrequency ablation in patients with nonvalvular atrial fibrillation: Insights from the China-AF cohort.

Heart rhythm·2026

Same author

Validation of Smartwatches Integrated With Photoplethysmography for Continuous Evaluation of Atrial Fibrillation Burden.

JACC. Clinical electrophysiology·2026

Same journal

Tracking Synthetic Adhesins on Bacterial Surfaces with Immunofluorescence Microscopy.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Post-Selection Methods for Analyzing mRNA Display Selections and Optimization of Hits.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

High-Performance Computing in Tandem Mass Spectrometry (MS/MS) Peptide Identification.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Engineering and Adapting Disulfide-Containing Proteins to Enable Intracellular Functionality.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

AI-Driven Protein Research: From Prediction to Design.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Methods for the In Vitro Selection of Protein and Peptide Libraries Using mRNA Display.

Methods in molecular biology (Clifton, N.J.)·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Apr 13, 2026

Identification of Protein Complexes in Escherichia coli using Sequential Peptide Affinity Purification in Combination with Tandem Mass Spectrometry

Identification of Protein Complexes in Escherichia coli using Sequential Peptide Affinity Purification in Combination with Tandem Mass Spectrometry

Published on: November 12, 2012

Repurposing Proximity-Dependent Protein Labeling (BioID2) for Protein Interaction Mapping in E. coli.

Tom Killelea¹, Fiona E Kemm², Liu He²

¹School of Life Sciences, University of Nottingham, Nottingham, UK. tom.killelea@nottingham.ac.uk.

Methods in Molecular Biology (Clifton, N.J.)

|August 15, 2024

Summary

This summary is machine-generated.

This study introduces a BioID2 proximity labeling toolkit for E. coli, enabling the study of protein-protein interactions. The method facilitates the identification of interaction networks within living cells.

Keywords:

BioID2 Biotinylation CheA Chemotaxis E. coli Protein-protein interaction Proximity-dependent labeling

More Related Videos

Inducible LAP-tagged Stable Cell Lines for Investigating Protein Function, Spatiotemporal Localization and Protein Interaction Networks

Inducible LAP-tagged Stable Cell Lines for Investigating Protein Function, Spatiotemporal Localization and Protein Interaction Networks

Published on: December 24, 2016

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

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

Published on: April 20, 2018

Related Experiment Videos

Last Updated: Apr 13, 2026

Identification of Protein Complexes in Escherichia coli using Sequential Peptide Affinity Purification in Combination with Tandem Mass Spectrometry

Identification of Protein Complexes in Escherichia coli using Sequential Peptide Affinity Purification in Combination with Tandem Mass Spectrometry

Published on: November 12, 2012

Inducible LAP-tagged Stable Cell Lines for Investigating Protein Function, Spatiotemporal Localization and Protein Interaction Networks

Inducible LAP-tagged Stable Cell Lines for Investigating Protein Function, Spatiotemporal Localization and Protein Interaction Networks

Published on: December 24, 2016

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

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

Published on: April 20, 2018

Area of Science:

Molecular Biology
Biochemistry
Systems Biology

Background:

Protein-protein interactions are crucial for cellular functions.
Proximity-dependent labeling methods, like BioID2, reveal these interactions in vivo.
Adapting BioID2 for E. coli requires specific optimization.

Purpose of the Study:

To develop and present a BioID2 toolkit tailored for E. coli.
To demonstrate the utility of this toolkit using the chemotaxis protein CheA.
To establish a methodology for identifying protein interaction networks in E. coli.

Main Methods:

Constructed plasmids with BioID2 fused to target proteins (e.g., CheA) at N or C termini.
Optimized fusion constructs with an 8×GGS linker for expression in E. coli.
Developed protocols for protein expression, in vivo biotinylation, and analysis of biotinylated interactors.

Main Results:

Successfully created and verified CheA-BioID2 fusion proteins in E. coli.
Demonstrated in vivo biotinylation of proteins in proximity to CheA-BioID2.
Established a workflow for capturing and identifying biotinylated protein interactors.

Conclusions:

The developed BioID2 toolkit is effective for studying protein-protein interactions in E. coli.
This methodology provides a valuable resource for mapping interaction networks in bacterial systems.
The toolkit enables the discovery of novel protein interactions relevant to biological mechanisms.