Title
Transcriptional coactivator protein p300. Kinetic characterization of its histone acetyltransferase activity
UMMS Affiliation
Department of Biochemistry and Molecular Pharmacology
Publication Date
2001-09-07
Document Type
Article
Subjects
Acetyltransferases; Amino Acid Sequence; Animals; Catalysis; Dose-Response Relationship, Drug; Histone Acetyltransferases; Histones; Humans; Kinetics; Models, Chemical; Molecular Sequence Data; Nuclear Proteins; Peptides; Plasmids; Protein Binding; *Saccharomyces cerevisiae Proteins; Sequence Homology, Amino Acid; Time Factors; Trans-Activators; *Transcription, Genetic; *Transcriptional Activation; Xenopus laevis
Disciplines
Biochemistry | Enzymes and Coenzymes | Medicinal-Pharmaceutical Chemistry | Therapeutics
Abstract
The p300/cAMP response element-binding protein-binding protein (CBP) family members include human p300 and cAMP response element-binding protein-binding protein, which are both important transcriptional coactivators and histone acetyltransferases. Although the role of these enzymes in transcriptional regulation has been extensively documented, the molecular mechanisms of p300 and CBP histone acetyltransferase catalysis are poorly understood. Herein, we describe the first detailed kinetic characterization of p300 using full-length purified recombinant enzyme. These studies have employed peptide substrates to systematically examine the substrate specificity requirements and the kinetic mechanism of this enzyme. The importance of nearby positively charged residues in lysine targeting was demonstrated. The strict structural requirement of the lysine side chain was shown. The catalytic mechanism of p300 was shown to follow a ping-pong kinetic pathway and viscosity experiments revealed that product release and/or a conformational change were likely rate-limiting in catalysis. Detailed analysis of the p300 selective inhibitor Lys-CoA showed that it exhibited slow, tight-binding kinetics.
DOI of Published Version
10.1074/jbc.M104736200
Source
J Biol Chem. 2001 Sep 7;276(36):33721-9. Link to article on publisher's site. Epub 2001 Jul 9.
Journal/Book/Conference Title
The Journal of biological chemistry
Related Resources
Repository Citation
Thompson, Paul R.; Kurooka, H.; Nakatani, Y.; and Cole, P. A., "Transcriptional coactivator protein p300. Kinetic characterization of its histone acetyltransferase activity" (2001). Thompson Lab Publications. 88.
https://escholarship.umassmed.edu/thompson/88
Comments
At the time of publication, Paul Thompson was not yet affiliated with UMass Medical School.