title: 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.
authors.affiliation: Department of Pharmacology and Molecular Sciences, Johns Hopkins School of Medicine, Baltimore, Maryland 21205, USA.
authors.forename: Kannan R
authors.initials: KR
authors.lastname: Karukurichi
authors.forename: Ling
authors.initials: L
authors.lastname: Wang
authors.forename: Lerna
authors.initials: L
authors.lastname: Uzasci
authors.forename: Cara Marie
authors.initials: CM
authors.lastname: Manlandro
authors.forename: Qing
authors.initials: Q
authors.lastname: Wang
authors.forename: Philip A
authors.initials: PA
authors.lastname: Cole
mesh_terms: D000105:Acetyl Coenzyme A / Q000378:metabolism; D000107:Acetylation; D000595:Amino Acid Sequence; D008958:Models, Molecular; D008969:Molecular Sequence Data; D050880:p300-CBP Transcription Factors / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D000099224:p300-CBP-Associated Factor
chemical_list: D000105:Acetyl Coenzyme A; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor