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

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.

SUBMITTER: Karukurichi KR

PROVIDER: S-EPMC2811879 | biostudies-literature | 2010 Feb

REPOSITORIES: biostudies-literature

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

Karukurichi Kannan R KR Wang Ling L Uzasci Lerna L Manlandro Cara Marie CM Wang Qing Q Cole Philip A PA

Journal of the American Chemical Society 20100201 4

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 ...[more]

PMID: 20063892

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Project description:This SuperSeries is composed of the SubSeries listed below.

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