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

PubChem ID

167551

Name

Anacardic Acid

Measurement

0.6μM - IC50

PubChem ID

5490884

Name

Garcinol

Measurement

0.34μM - IC50

PubChem ID

25193530

Name

CPTH2

Measurement

52μM - IC50

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei