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

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

Author information

1
Department of Pharmacology and Molecular Sciences, Johns Hopkins School of Medicine, Baltimore, Maryland 21205, USA.

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.

PMID:
20063892
PMCID:
PMC2811879
DOI:
10.1021/ja909466d
[Indexed for MEDLINE]
Free PMC Article

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