You are interested in understanding the mechanism of action of your favorite mou

Question

You are interested in understanding the mechanism of action of your favorite mouse protein (YFP). You suspect that YFP interacts with a histone demethylase, KDM1. Assuming you have all necessary reagents, describe an experimental strategy to determine the interaction of YFP with KDM1. Include all necessary reagents, expected results etc. to clarify your answer. You suspect that YFP and KDM1 are part of a large multi-protein complex. Briefly describe an experimental approach you would use to identify all the proteins of this complex.

Explanation / Answer

(a) the technique that can be used to study the interaction is by FRAP ie flouresnce recovery after phto bleaching..

In this technique the mobile protein of interst ie mouse YFP is tagged with a GFP (green flouresnce protein). Then is it allowed to enter into the body whose position can be traced by the flouresnce microscope.. When this protein is bound to the receptor ie KDM1. As the principle of FRAP the flouresnce from the YFP Protein is bleached by the binding of KDM1 which can very tracked by the microsope.. Hence it can also show the evidence of binding of these two..

(b) in order to determine the protein complex we can use NMR (nuclear magnetic resonance) spectroscopy. In this technique we can find out the all the subunits present in the protein..!!

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