TurboID Service

What Is TurboID?

TurboID is an engineered variant of the bacterial biotin ligase BirA, developed through directed evolution to enable rapid proximity-dependent protein labeling. Following biotin labeling, biotinylated proteins are enriched through streptavidin affinity capture and subsequently identified using LC-MS/MS.

Hollstein, L. S. et al. Scientific Reports. 2022.

Figure 1. Workflow of a TurboID Experiment Using SCI1 as Bait

The rapid labeling capability of TurboID makes it suitable for identifying proteins involved in weak or transient associations that may be lost during conventional affinity purification. It also enables researchers to profile the local protein environment surrounding a target protein. When combined with appropriate experimental controls and quantitative analysis, TurboID can be used to compare changes in protein proximity across biological conditions, identify candidate interaction partners, and support studies of dynamic protein networks and subsequent functional validation.

 

When TurboID Is a Good Fit

1. Weak or Transient Protein Associations

Identify candidate proteins in the vicinity of a target protein that may be lost during conventional affinity purification.

 

2. Rapid Biological Events

Use an approximately 10-minute labeling window to investigate protein changes associated with stimulus responses or dynamic cellular processes.

 

3. Subcellular Protein Mapping

Identify candidate proteins surrounding a target protein within specific subcellular compartments.

 

4. Comparison Across Experimental Conditions

Compare target-proximal protein profiles across treatments or biological states to identify differentially enriched candidate proteins.

TurboID vs miniTurbo vs APEX/APEX2: How to Choose?

 

Comparison	TurboID	miniTurbo	APEX/APEX2
Labeling Mechanism	Biotin ligase-mediated proximity labeling	Biotin ligase-mediated proximity labeling	Peroxidase-mediated proximity labeling
Approximate Labeling Time	~10 min	~30-60 min	~1 min
Approximate Labeling Radius	~10 nm	Depends on the experimental system	~20 nm
Key Advantage	Rapid labeling for profiling target-proximal proteins	Alternative biotin ligase system for projects suited to its labeling characteristics	Ultrafast labeling for capturing rapid biological events
Best-Fit Research Needs	Weak or transient protein associations and local protein profiling	Protein neighborhood profiling when a longer labeling window is appropriate	Rapid signaling events and dynamic changes in local protein environments
Key Considerations	Labeling conditions and target localization affect protein coverage	Labeling efficiency and background require experimental assessment	Requires peroxidase substrates and controlled reaction conditions

Note: Labeling times and spatial ranges are approximate and depend on the experimental system, labeling conditions, and analytical approach.

Service at MtoZ Biolabs

MtoZ Biolabs combines TurboID proximity labeling, protein enrichment, and LC-MS/MS analysis to provide customized services for investigating protein proximity and local protein environments.

 

1. Rapid Proximity Labeling in Living Cells

Use TurboID to achieve rapid labeling in approximately 10 minutes, enabling the identification of proteins near a target under specific biological conditions.

 

2. Analysis of Weak or Transient Protein Associations

Identify candidate proximal proteins that may be difficult to retain using conventional AP-MS, providing insights into dynamic protein associations.

 

3. Subcellular Proximity Proteomics

Identify proteins surrounding a target protein within specific subcellular compartments to characterize its local protein environment.

 

4. Comparative Proximity Proteomics

Compare target-proximal protein profiles across experimental conditions to identify differentially enriched candidate proteins.

 

Analysis Workflow

1. Fusion Construct Development

Generate an expression construct encoding the target protein fused to TurboID.

 

2. Cell Culture and Target Protein Expression

Establish the appropriate cellular system through transient transfection or stable expression, depending on project requirements.

 

3. Biotin Labeling

Label proteins in the vicinity of the target protein under selected experimental conditions.

 

4. Western Blot Analysis

Evaluate protein biotinylation using Western blotting.

 

5. Streptavidin Enrichment

Enrich biotinylated proteins through streptavidin affinity capture.

 

6. LC-MS/MS Analysis

Analyze enriched proteins using high-resolution Orbitrap mass spectrometry.

 

7. Database Searching and Data Analysis

Search acquired mass spectrometry data against an appropriate protein reference database and summarize the identification results.

Why Choose MtoZ Biolabs?

1. Extensive experience in MS-based proteomics.

2. Expertise in protein-protein interaction analysis and proximity labeling techniques.

3. Optimized TurboID workflows validated across multiple experimental systems.

4. High-resolution LC-MS/MS platforms for sensitive protein detection.

5. Flexible project customization based on specific research objectives.

Experimental Instruments

1. Mass Spectrometry Platform

High-resolution Orbitrap mass spectrometers.

2. Data Acquisition Modes

DDA or DIA, selected according to project requirements.

3. Data Analysis Software

MaxQuant, DIA-NN, or PEAKS, depending on the acquisition mode and analytical requirements.

4. Protein Reference Database

UniProt or a custom database provided by the client.

Applications

1. Identification of transient or stimulus-dependent protein interactions.

2. Mapping of local protein environments within subcellular compartments.

3. Investigation of dynamic changes in signaling pathways.

4. Characterization of membrane protein-associated networks.

5. Investigation of protein organization under changing cellular conditions.

6. Comparative analysis of protein interaction landscapes across treatments or time points.

Sample Submission Requirements

Item	Requirements
Sample Type	Cells expressing the target protein–TurboID fusion construct through transient transfection or stable expression, with information on the target protein and construct status
Recommended Sample Amount	At least 2 × 10⁷ cells per sample
Minimum Sample Amount	1 × 10⁷ cells per sample
Sample Preparation	Submit frozen cell samples
Storage and Shipping	Ship frozen cell samples on dry ice
Experimental Information	Provide the target protein, construct status, labeling status, experimental groups, and available control information

The recommended and minimum cell amounts apply to each individual sample. Before shipping, please confirm starting material and sample preparation requirements with our technical team, particularly for projects involving fusion construct development and live-cell labeling.

Deliverables

1. Raw Mass Spectrometry Data

Original mass spectrometry data files.

2. Protein Identification Results

Excel files containing identified proteins and available supporting identification evidence.

3. Basic PDF Analysis Report

A report summarizing the experimental workflow and protein identification results.

4. Bioinformatics Analysis

GO enrichment, KEGG, and protein-protein interaction network analysis can be performed based on project requirements.

 

FAQs

Q1: When should APEX/APEX2 be considered instead of TurboID?

APEX/APEX2 may be considered when a study requires a different labeling time window or spatial range, including certain investigations of proteins associated with metabolic pathways. The appropriate method should be selected based on the target protein, biological system, and research objectives.

 

Contact Us

Share your target protein, biological system, research objectives, fusion construct status, experimental groups, available controls, and sample information with MtoZ Biolabs. Our technical team can use this information to evaluate project feasibility and determine an appropriate TurboID proximity labeling and LC-MS/MS analysis strategy.