2 You have identified two proteins that are believed to inte

2) You have identified two proteins that are believed to interact and bind together. The exact binding site has not been identified. However, it is believed to be at the carboxyl end of one protein to the amino end of the other protein. How could you verify this interaction and locate the binding sites?

Solution

The building blocks of proteins are amino acids, which are small organic molecules that consist of an alpha (central) carbon atom linked to an amino group, a carboxyl group, a hydrogen atom, and a variable component called a side chain

Co immuno precipitation is the best method to analyse the interaction because while doing the procedure,it can identify other proteins it directly or indirectly interacts with the core protein or protein of interest.

Y2H is a more specific test. Here, the DNA binding domain and the activation domain are split up. Using these halves you can investigate whether two proteins of interest bind. The DNA binding domain is fused to one protein of interest (bait), and the activation domain is fused to another protein of interest (prey). If the two proteins of interest (bait and prey) bind, then you \"reunite\" the activation domain and DNA binding domain. As a result you will get transcription of a downstream factor.

Chromatin immunoprecipitation tests can also used to find whether a protein interacts with DNA. In my opinion, Western blot can be used prior to testing to ensure the presence of protein.