Activity-based substrate profiling for Gcn5-related N-acetyltransferases: The use of chloroacetyl-coenzyme A to identify protein substrates

Michael Yu, Luiz Pedro Sorio De Carvalho, Guangxing Sun, John S. Blanchard

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

The Gcn5-related N-acetyltransferases (GNAT) comprise one of the largest enzyme superfamilies, with over 10000 known members represented in all kingdoms of life. ChloroacetylCoenzymeA was prepared and demonstrated to be a substrate for several GNAT members. ChloroacetylCoA (ClAcCoA) is used by the Hat1 histone acetyltransferase to correctly acetylate histone H4 in a mixture of histone proteins. Chloroacetylation can be assessed by the subsequent reaction of the chloroacetylated product with thiol-containing compounds, including those with fluorescent or affinity (His8) tags. The bacterial RimL N-acetyltransferase also uses ClAcCoA to chloroacetyl the α-amino group of its cognate substrate, the ribosomal L12 protein, and this reaction can be observed in crude extracts. ChloroacetylCoA is a reagent that can be used to identify the unknown substrate(s) for this large family of functionally uncharacterized enzymes.

Original languageEnglish (US)
Pages (from-to)15356-15357
Number of pages2
JournalJournal of the American Chemical Society
Volume128
Issue number48
DOIs
Publication statusPublished - Dec 6 2006

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ASJC Scopus subject areas

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

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