Abstract
Fluorescence spectroscopy is routinely used for the determination of the interaction of a ligand with a protein. The quick detection of the interaction between the ligand and the protein is one of the most significant advantages of fluorescence spectroscopic methods. In this chapter, we have described assays to monitor drug –tubulin interactions using several fluorescence spectroscopic techniques. We have provided detailed protocols for different assays for investigating tubulin–drug interactions with key practical considerations for performing the experiments. We have also discussed how to deduce the binding parameters by fitting the fluorescence change data in different binding isotherms. Further, we have described detailed protocols to monitor the binding site of a ligand on tubulin by competitive inhibition. Though the methods are described for tubulin, these methods can also be used to monitor any drug –protein interactions.
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Abbreviations
- DMSO:
-
Dimethyl sulfoxide
- EGTA:
-
Ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid
- GTP:
-
Guanosine triphosphate
- MgCl2:
-
Magnesium chloride
- PIPES :
-
Piperazine-N,N′-bis(2-ethanesulfonic acid)
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Acknowledgments
DP thanks JC Bose fellowship (JCB/2019/000016) from the Department of Science and Technology, Government of India.
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Kumari, A., Panda, D. (2022). Investigating Tubulin-Drug Interaction Using Fluorescence Spectroscopy. In: Inaba, H. (eds) Microtubules. Methods in Molecular Biology, vol 2430. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1983-4_17
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DOI: https://doi.org/10.1007/978-1-0716-1983-4_17
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