Due to the effort of many scientists, we can conclude many kinds of methods to be used in protein and protein interaction experiment. They are IP, Two-Hybrid System, FRET and BRET. Here I will introduce some methods of this experiment.
Detection of protein and protein interaction
1. Yeast two-hybrid systerm
The yeast two-hybrid system is now widely used in the protein and protein interaction experiment. The theory of this technology is that after target proteins combine with bait proteins. The bait protein binds to the promoter of the reporter genes. And then the reporter genes will start to express in yeast cells. If the expression products are detected, we can conclude the two proteins have interaction.
2. Phage display technology
What we should do is to link a monoclonal antibody of DNA sequences to the protein of the phage, the corresponding monoclonal antibody will express on the surface while the phage is growing. And then we can pass the phage through the column. If there are target proteins, it will combine with the corresponding antibodies. This is called phage display technology.
3. Plasmon resonance technology
Plasmon resonance technology, especially the Surface Plasmon Resonance (SPR) has become the new method for the protein and protein interaction field. The theory of this technology is that we adsorb a nanoscale film to the bait protein. The feature of the resonance films will be changed after the target proteins combine with the bait protein. Therefore, we can analyze if the two proteins have interaction. The advantages of SPR technology are lying in no need of markers or dye, the process of interaction can be observed.
4. Fluorescence energy transfer
Fluorescence energy transfer is widely used in the distance and interaction between moleculars. When used with the help of fluorescence microscopy, FRET technology can obtain the information of proteins, fat, DNA and RNA in creatures. As the development of GFP, the FRET fluorescence microscopy may help people achieve the detection of interaction of moleculars in creatures’ bodies.
5. Antibody and protein array technology
The coming of protein array has pave a new way for the researches of proteomics. As we all know that the main research point for proteomics is the quantitative of protein levels in different physiological states. The protein array is the best tool for this field among many arrays. Of course, these arrays have been used in the clinical analysis.
6.Co-immunoprecipitation
This technology is also used to study the interaction between proteins. The first step is to add the special antibody in cell lysates. And then we can add the SPA, which can be binded with antibodies, after the incubation.
As we mentined above, there are many kinds of technologies in protein and protein interaction field. However, we can combine them to find the best and the most useful method with the help of current technologies.