Analysis of p300/CBP histone acetyltransferase regulation using circular permutation and semisynthesis

J Am Chem Soc. 2010 Feb 3;132(4):1222-3. doi: 10.1021/ja909466d.

Abstract

The histone acetyltransferase (HAT) p300/CBP has been shown to undergo autoacetylation on lysines in an apparent regulatory loop that stimulates HAT activity. Here we have developed a strategy to introduce acetyl-Lys at up to six known modification sites in p300/CBP HAT using a combination of circular permutation and expressed protein ligation. We show that these semisynthetic, circularly permuted acetylated proteins retain high affinity for an acetyl-CoA substrate analogue and that HAT activity correlates positively with degree of acetylation. This study provides novel evidence for control of p300/CBP HAT activity by site-specific autoacetylation and outlines a potentially general strategy for using expressed protein ligation and circular permutation to chemically interrogate internal regions of proteins.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Acetyl Coenzyme A / metabolism
  • Acetylation
  • Amino Acid Sequence
  • Models, Molecular
  • Molecular Sequence Data
  • p300-CBP Transcription Factors / chemical synthesis
  • p300-CBP Transcription Factors / chemistry*
  • p300-CBP Transcription Factors / metabolism*

Substances

  • Acetyl Coenzyme A
  • p300-CBP Transcription Factors
  • p300-CBP-associated factor