Hello everyone,
I would like to understand the interaction between my drug of interest to a protein (known). Basically, I want to study how strong the interaction is. Is there any way to measure it?
Thanks in advance,
Arun.
Arun Pandian Chandrasekaran , You can try fluorescence based analysis if you can label either your drug or the protein with a fluorescent label. Please refer to "Click chemistry approach"
There are various methods to study the protein drug interaction. If you have purified protein in sufficient amount (in mg quantity) you can use isothermal titration calorimetry which gives information regarding enthalpy, entropy and kd value for binding. Further it gives detailed information regarding the type of interaction ionic or non ionic and also molecular interaction phenomenon such as enthalpy entropy compensation. If you are interested only in the binding constants koff and kon you can use Biolayer interferometery.
Hope this helps
Well, it depends on your protein and the drug candidate. ITC is good provided both the protein and the ligand are in the same solvent. If the protein is in aqueous buffer system and the ligand is in organic solvent, you need to make sure that too much heat is not released when you add just the organic solvent (without ligand as control experiment) to the protein. If it does, it will mask the small change that will happen when your ligand is present. ITC doesn't work for protein-ligand systems in my lab for the above reasons.
Fluorescence is a great method if you have Tryptophans. You may be able to monitor Trp fluorescence (if acccessible to ligand or there is change in conformation that affects Trp environment) as you add the ligand.
You can also use circular dischorisim (CD) as well monitoring either 180-260 nm region. You can also use 2D 1H-15N-HSQC if you have N-15 labeled protein samples. SPR is the other method at your disposal.
Thank you, everyone, for your valuable suggestions. I will look at each one them in detail.
Hi Arun,
As mentioned in the previous comments, there are many reliable techniques for measuring drug-protein interactions. If you don’t have the resources and equipment, then you can work together with a reliable partner to solve this problem. I know that Creative Biostructure has drug discovery-related services. This is a very professional structural biology company. I think it is very good, you can try it.
Hope it helps!
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