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Fluorogenic Assays for the Defatty-Acylase Activity of Sirtuins

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Protein Lipidation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2009))

Abstract

Sirtuins are type III histone deacetylases (HDAC) that uses nicotinamide adenine dinucleotide as cosubstrate. Dysfunction of sirtuins is implicated in wide varieties of human diseases. As such, there has been increased interest in the development of small molecule to modulate sirtuin activities. Besides deacetylase activity, recent studies suggest SIRT1, 2, 3, and 6 efficiently remove fatty acyl groups on lysine. In vitro sirtuin enzymatic activity assays established so far are mainly based on the deacetylation activity. Here, we describe a fluorogenic assay for monitoring defatty-acylase activity of SIRT1, 2, 3 and 6 using peptide substrates. This assay can be utilized to evaluate sirtuin modulators in high-throughput manners.

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Correspondence to Hening Lin .

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Young Hong, J., Cao, J., Lin, H. (2019). Fluorogenic Assays for the Defatty-Acylase Activity of Sirtuins. In: Linder, M. (eds) Protein Lipidation. Methods in Molecular Biology, vol 2009. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9532-5_10

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  • DOI: https://doi.org/10.1007/978-1-4939-9532-5_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9531-8

  • Online ISBN: 978-1-4939-9532-5

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