Transcriptional coactivator protein p300. Kinetic characterization of its histone acetyltransferase activity
UMass Chan Affiliations
Department of Biochemistry and Molecular PharmacologyDocument Type
Journal ArticlePublication Date
2001-09-07Keywords
AcetyltransferasesAmino 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
Biochemistry
Enzymes and Coenzymes
Medicinal-Pharmaceutical Chemistry
Therapeutics
Metadata
Show full item recordAbstract
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.Source
J Biol Chem. 2001 Sep 7;276(36):33721-9. Link to article on publisher's site. Epub 2001 Jul 9.DOI
10.1074/jbc.M104736200Permanent Link to this Item
http://hdl.handle.net/20.500.14038/50083Notes
At the time of publication, Paul Thompson was not yet affiliated with UMass Medical School.
Related Resources
Link to Article in PubMedae974a485f413a2113503eed53cd6c53
10.1074/jbc.M104736200