Identification of Protein-Protein Interaction Networks in Cells

Protein-protein interactions (PPIs) constitute an indispensable core component of cellular life activities, spanning multiple temporal and spatial dimensions and profoundly influencing crucial cellular processes such as cell cycle regulation, protein synthesis and secretion, signal transduction, and metabolism. Therefore, a deep investigation into protein interactions plays a pivotal role in uncovering the mysteries of molecular regulatory networks.

Currently, commonly used techniques for studying protein interactions include co-immunoprecipitation, pull-down assays, protein microarrays, and others. However, these methods face numerous challenges in practical applications, such as excessive dependence on antibodies and difficulty in capturing transient or weak interactions effectively. Proximity labeling technology has emerged to address these challenges, whereby enzymes biotinylate target proteins in close proximity, followed by affinity purification and mass spectrometry characterization of PPIs. Among these, proximity labeling methods based on ascorbate peroxidase (APEX) and biotin ligase are the most widely used for PPI identification. TurboID, a novel technology platform developed based on proximity-dependent biotin identification (BioID), employs a mutated form of E. coli biotin ligase. CoLinkBio's mass spectrometry-based proximity labeling technology platform encompasses various techniques including BioID, APEX, TurboID, and others. These techniques demonstrate outstanding applicability in live cell systems, particularly in capturing transient or dynamic protein interaction events, as well as interactions involving low-abundance proteins or hydrophobic membrane proteins.

Technical Platform

Taking the TurboID technology platform as an example, this platform fuses a mutated biotin ligase to the C-terminus of the target protein. In overexpressing cells, the addition of biotin stimulus enables rapid biotinylation of interacting proteins of the target protein within 10 minutes. This labeling process is not only fast and low-background but can also be smoothly conducted even at room temperature. Consequently, the TurboID technology platform significantly expands its applicability, being suitable not only for mammalian cell systems but also for various biological systems such as plants and insects. Subsequently, by isolating and enriching biotin-labeled proteins followed by enzymatic digestion, peptide samples can be easily obtained for mass spectrometry analysis, enabling in-depth investigation of protein interactions and their functions.

Our Advantages

1. Professional Excellence: Our team boasts extensive experience and publications in top journals, offering industry-leading technical services.
2. Efficient Solutions: We employ reliable methods to drive projects forward swiftly, providing worry-free solutions.
3. Rigorous Quality Management: Adhering to ISO 9001 standards, our mature quality management system ensures the authenticity and reliability of our reports.
4. Systematic Project Management: From consultation to report delivery, we provide timely progress updates, ensuring customer satisfaction and efficient project execution.
5. Cutting-Edge Equipment: Equipped with advanced mass spectrometers like the Thermo Fisher Orbitrap Exploris 480 and Bruker timsTOF, we facilitate groundbreaking research.

Our Service

Project	Identification of Protein-Protein Interaction Networks in Cells
Sample	Cell lysate, live cells
Hardware Platform	Non-contact ultrasonic cell pulverizer，ChemiDoc MP Imaging System，Orbitrap Fusion Lumos Tribrid/Orbitrap Exploris 480/Q Exactive HF-X/timsTOF Pro 2 mass spectrometer
Project Duration	4-6 weeks
Deliverables	Project Report (including experimental procedures, data analysis charts, bioinformatics analysis results)
Price	Click to consult

Case Study

Project Aim: Analyze changes in the protein interactome of the target protein (POI) in cells before and after drug treatment.
Solution: Utilize a proximity labeling strategy by constructing a POI-TurboID expression vector, overexpressing it in cells, and setting up drug-treated and control groups for biotinylation. Enrich the protein interactome in cells, followed by qualitative and quantitative analysis of differences in biotinylated proteins to characterize changes in the POI interactome.
Data visualization: