Abstract
Among the different posttranslational modifications (PTMs) that significantly regulate the protein function, lysine acetylation has become the major focus, especially to understand the epigenetic role of the acetyltransferases, in cellular physiology. Furthermore, dysfunction of these acetyltransferases is well documented under pathophysiological conditions. Therefore, it is important to understand the dynamic structure–function relationship of acetyltransferases in a relatively less complicated and faster method, which could be efficiently exploited to design and synthesis of small molecule modulators (activators/inhibitors) of these enzymes for in vivo functional analysis and therapeutic purposes. We have developed surface-enhanced Raman scattering (SERS) method, for acetyltransferases towards this goal. By employing SERS, we have not only demonstrated the autoacetylation induced structural changes of p300 enzyme but also could use this technique to characterize and design potent, specific inhibitors as well as activators of the p300. In this chapter we shall describe the methods in detail which could be highly useful for other classes of HATs and PTM enzymes.
Authors Mohammed Arif, Dhanasekaran Karthigeyan, and Soumik Siddhanta contributed equally.
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Acknowledgment
We would like to thank JNCASR, Department of Science and Technology (DST), Govt. of India, and Chromatin and Disease Programme Support, Department of Biotechnology (DBT), Govt. of India, for funding. DK and SS would like to thank the Council for Scientific and Industrial Research for the Senior Research Fellowship. TKK is a recipient of Sir JC Bose National Fellowship, DST.
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Arif, M., Karthigeyan, D., Siddhanta, S., Kumar, G.V.P., Narayana, C., Kundu, T.K. (2013). Analysis of Protein Acetyltransferase Structure–Function Relation by Surface-Enhanced Raman Scattering (SERS): A Tool to Screen and Characterize Small Molecule Modulators. In: Hake, S., Janzen, C. (eds) Protein Acetylation. Methods in Molecular Biology, vol 981. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-305-3_19
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DOI: https://doi.org/10.1007/978-1-62703-305-3_19
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