Biological Information
Background Information:
Histone acetyltransferases (HAT) are enzymes that covalently modify the N-terminal lysine of all four core histones in nucleosomes by the addition of acetyl groups from acetyl-CoA. HAT enzymes regulate transcription via chromatin remodeling; HAT’s are recruited on to the chromatin template and transcription is enhanced by acetylation of promoter proximal nucleosomal histones. Upon acetylation the histone complex relaxes facilitating other proteins to act on the DNA. P300 acetylase activity is not limited to histones since it can also acetylate non histone targets like p53 and ERa. P300 directly interacts with transcription factors and acts as a global transcriptional coactivator; it plays a critical role in a variety of cellular process including cell cycle control, differentiation, and apoptosis. P300 deregulation is believed to play an important role in Cancer.
Family:
Epigenetic
Sub Family:
Histone Acetyltransferase (HAT)
Class:
Writer
Protein Name:
P300
Uniprot Number:
Q09472
Protein Aliases:
histone acetyltransferase p300
Gene Name:
EP300
Gene ID:
2033
Gene Aliases:
p300|KAT3B
Accession Number:
NM_001429
Organism:
Human
Construct Details:
Fragment (1284-1673), Recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Radiometry
Measured Response:
Scintillation
Testing Information
Procedure Summary:
Human recombinant histone acetyltransferase P300 expressed in E. coli. is used. Test compound and/or vehicle is preincubated with 1 μg/ml enzyme‡ in modified Tris-HCl buffer pH 8.5 for 90 minutes at 37°C. The reaction is initiated by adding 3.33 μg/ml Core Histone and 0.1 μCi [3H]Acetyl-CoA. Separate bound and free ligand by filtermats and count to determine the amount of [3H]Acetyl histone formed. Compounds are screened at 10 μM.
Substrate:
3.33 µg/ml Histone
Incubation:
90 min at 37°C
Control Inhibitor:
Garcinol
Criteria For Significance:
≥ 50% of max stimulation or inhibition
Test Concentration / Dose:
Suggested concentration for initial testing is 10 uM
Test Sample Requirements:
Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).
Minimum Order Quantity:
10
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei