Here I am using two different wavelengths to target different moiety in BSA by fluorescence quenching method.
Where I kept BSA as fluorescence active and drug (fluorescence inactive). Now I am studying interaction of a drug with its differential concentration by keeping BSA concentration fix.
So, from this observation can I find or assume possible binding site in BSA for that drug ?
When you excite BSA with two different WL in fact you are selecting the intrinsic probe. If only triptophan or tyrosin. Yes, you can titrate the drug against BSA, keeping fixed the concentration, and using the Ster-Volmer graphs and equations you can infere about binding. See my publications on the matter specially those from 2004.
There are others methods to study ligand binding to proteins and I recommend reading Dr. Ulrich Kraag-Hansen works.
Thank you Sir.Yes i am following this method.I have targeted triptophan & tyrosin residues by 280 & 293nm.I have did this work with two or more drug.
But towards writing paper,it seems to me that Is only fluorescence quenching analysis is enough for this study ?because i have seen some of recent papers where they follow other method along with fluorescence like Circular dicromism, FTIR,UV etc.
Sir,it will be convenient for me if you suggest me a journal name,where i can submit my paper.
Thank you.
Yes, there is a tendency to join methods to better conclude, and CD is one of them because you can monitor changes in spatial arrange of the protein along the titration.
About journals, if your focus is more driven to methodology I suggest Spectrochimica Acta B, But if the focus is in results you can submit to Journal of Photochemistry and Photobiiology.
Yes, you can study drug binding with fluorescence quenching method by titrating a fixed amount of the protein with increasing concentration of the drug. Then you can use different linear transformations to get the value of binding constant. However, these methods have their own limitations. But many people are still using these methods.
Important thing to remember to do inner filter effect correction in the fluorescence data.
Simply getting the binding constant and number of binding site will not be sufficient to get a publication. Show the type of quenching (dynamic or static) involved which can be done by carrying out this titration at three different temperatures.
Characterization of the binding site on BSA can be made by using marker ligands for Sudlow's site I and II.
Best.
To determine the values of the binding constant and the number of binding sites dyes with BSA, the titration was performed by the method of Scatchard. We present the results of a study on the relationship between the structure and binding of dye and its derivatives with BSA in the paper in the jornal Photonics and Optoelectronics volume 3 issue 1,January 2014.
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