Azalysine analogues as probes for protein lysine deacetylation and demethylation

Blair C.R. Dancy, Shonoi A. Ming, Romeo Papazyan, Christine A. Jelinek, Ananya Majumdar, Yan Sun, Beverley M. Dancy, William J. Drury, Robert J. Cotter, Sean D. Taverna, Philip A. Cole

Research output: Contribution to journalArticle

Abstract

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′-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.

Original languageEnglish (US)
Pages (from-to)5138-5148
Number of pages11
JournalJournal of the American Chemical Society
Volume134
Issue number11
DOIs
StatePublished - Mar 21 2012

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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