Azalysine Analogues as Probes for Protein Lysine Deacetylation and Demethylation

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• 作者：Blair C. R. Dancy ; Shonoi A. Ming ; Romeo Papazyan ; Christine A. Jelinek ; Ananya Majumdar ; Yan Sun ; Beverley M. Dancy ; William J. Drury ; III ; Robert J. Cotter ; Sean D. Taverna ; Philip A. Cole
• 刊名：The Journal of the American Chemical Society
• 出版年：2012
• 出版时间：March 21, 2012
• 年：2012
• 卷：134
• 期：11
• 页码：5138-5148
• 全文大小：532K
• 年卷期：v.134,no.11(March 21, 2012)
• ISSN：1520-5126

文摘

Reversible lysine acetylation and methylation regulate the function of a wide variety of proteins, including histones. Here, we have synthesized azalysine-containing peptides in acetylated and unacetylated forms as chemical probes of the histone deacetylases (HDAC8, Sir2Tm, and SIRT1) and the histone demethylase, LSD1. We have shown that the acetyl-azalysine modification is a fairly efficient substrate for the sirtuins, but a weaker substrate for HDAC8, a classical HDAC. In addition to deacetylation by sirtuins, the acetyl-azalysine analogue generates a novel ADP-ribose adduct that was characterized by mass spectrometry, Western blot analysis, and nuclear magnetic resonance spectroscopy. This peptide-ADP-ribose adduct is proposed to correspond to a derailed reaction intermediate, providing unique evidence for the direct 2鈥?hydroxyl attack on the O-alkylimidate intermediate that is formed in the course of sirtuin catalyzed deacetylation. An unacetylated azalysine-containing H3 peptide proved to be a potent inhibitor of the LSD1 demethylase, forming an FAD adduct characteristic of previously reported related structures, providing a new chemical probe for mechanistic analysis.