Application of Label Transfer in Protein Interaction Studies

The core mechanism of the label transfer technique involves using specific labeling reagents to link a target protein with interacting proteins, thereby enabling precise protein interaction analysis. 

 

1. Selection and Binding of Labeling Reagents

First, suitable labeling reagents are chosen, which typically possess photosensitive or chemically reactive properties capable of forming covalent bonds with the bait protein. After the protein complex forms, these labeling reagents are activated by light or chemical reactions, binding to the captured prey proteins and thereby labeling the proteins that interact with the target protein.

 

2. Initiation of Crosslinking Reaction

Once the protein complex is formed and labeled, the crosslinking reaction is initiated by UV irradiation or chemical treatment. The reactive groups in the crosslinker form covalent bonds with specific amino acid residues in the prey protein, stabilizing these transient protein interactions. The precision and efficiency of this step directly determine the outcome of the interaction analysis.

 

3. Label Transfer and Protein Separation

After crosslinking, the spacer arm of the crosslinker is cleaved chemically, releasing the labeled prey proteins. These labeled prey proteins can then be separated through radioactive detection, fluorescence analysis, or other affinity capture techniques, ultimately identifying the specific proteins interacting with the bait protein.

 

4. Detection and Analysis of Proteins

The labeled prey proteins are then detected and analyzed using mass spectrometry or Western blotting. These analytical methods are efficient in identifying proteins that interact with the target protein and further explore their roles in biological functions.

 

Applications of Label Transfer in Protein Interaction Studies

Firstly, it effectively captures weak and transient protein interactions, addressing the limitations of traditional methods. Secondly, this technique is particularly effective in studying complex protein networks, allowing for precise analysis of individual components within protein complexes. Additionally, label transfer offers new perspectives for studying specific protein modifications and functions, such as exploring key protein interactions in signal transduction pathways.

 

MtoZ Biolabs provides integrate label transfer protein interaction analysis service.