Product nameKAT3B/p300 Inhibitor Screening Assay Kit (Fluorometric)
Sample typeInhibitor compounds
Assay typeEnzyme activity
KAT3B/p300 Inhibitor Screening Assay Kit (Fluorometric) (ab196996) is an assay where histone H3 peptide and Acetyl CoA are used as acetylation substrates. KAT3B/p300 acetylates the Histone H3 peptide and generates Coenzyme A with a free thiol group (CoA-SH). CoA-SH reacts subsequently with a Thiol Detecting Probe in order to increase the amount of fluorescence in the solution, which can be measured at Ex/Em = 392/482 nm. In the presence of KAT2B/p300 specific inhibitors, the enzymatic activity is reduced ot completely abolished resulting in decreased or total loss of fluorescence.
This assay kit is a simple, sensitive and rapid tool to screen potential p300 inhibitors.
p300 or KAT3B, a histone acetyltransferase, contributes to transcriptional activation by acetylating chromatin on the lysine residues of H3 and H4 histones. There is growing evidence that p300 plays an important role in cancer cell proliferation and differentiation. p300 inhibitors have potential applications in cancer therapy.
KTA3B (K-(lysine) acetyltransferase 3B, EC 220.127.116.11), also known as p300 possesses intrinsic histone acetyltransferase (HAT) activity and is able to acetylate lysine residues present in both histone H3 and histone H4.
There is growing evidence taht p300 plays an important role in cancer cell proliferation and differentiation. p300 inhibitors have potential applications in cancer therapy.
Storage instructionsStore at -80°C. Please refer to protocols.
Components 100 tests Acetyl CoA 1 vial H3 Peptide 1 vial p300 Assay Buffer 1 x 20ml p300 Enzyme 1 x 0.1ml p300 Inhibitor 1 x 10µl Thiol Detecting Probe 1 x 0.2ml
FunctionFunctions as histone acetyltransferase and regulates transcription via chromatin remodeling. Acetylates all four core histones in nucleosomes. Histone acetylation gives an epigenetic tag for transcriptional activation. Mediates cAMP-gene regulation by binding specifically to phosphorylated CREB protein. Mediates acetylation of histone H3 at 'Lys-122' (H3K122ac), a modification that localizes at the surface of the histone octamer and stimulates transcription, possibly by promoting nucleosome instability. Also functions as acetyltransferase for nonhistone targets. Acetylates 'Lys-131' of ALX1 and acts as its coactivator in the presence of CREBBP. Acetylates SIRT2 and is proposed to indirectly increase the transcriptional activity of TP53 through acetylation and subsequent attenuation of SIRT2 deacetylase function. Acetylates HDAC1 leading to its inactivation and modulation of transcription. Acts as a TFAP2A-mediated transcriptional coactivator in presence of CITED2. Plays a role as a coactivator of NEUROD1-dependent transcription of the secretin and p21 genes and controls terminal differentiation of cells in the intestinal epithelium. Promotes cardiac myocyte enlargement. Can also mediate transcriptional repression. Binds to and may be involved in the transforming capacity of the adenovirus E1A protein. In case of HIV-1 infection, it is recruited by the viral protein Tat. Regulates Tat's transactivating activity and may help inducing chromatin remodeling of proviral genes. Acetylates FOXO1 and enhances its transcriptional activity.
Involvement in diseaseDefects in EP300 may play a role in epithelial cancer.
Chromosomal aberrations involving EP300 may be a cause of acute myeloid leukemias. Translocation t(8;22)(p11;q13) with KAT6A.
Rubinstein-Taybi syndrome 2
Sequence similaritiesContains 1 bromo domain.
Contains 1 KIX domain.
Contains 2 TAZ-type zinc fingers.
Contains 1 ZZ-type zinc finger.
DomainThe CRD1 domain (cell cycle regulatory domain 1) mediates transcriptional repression of a subset of p300 responsive genes; it can be de-repressed by CDKN1A/p21WAF1 at least at some promoters. It conatins sumoylation and acetylation sites and the same lysine residues may be targeted for the respective modifications. It is proposed that deacetylation by SIRT1 allows sumoylation leading to suppressed activity.
modificationsAcetylated on Lys at up to 17 positions by intermolecular autocatalysis. Deacetylated in the transcriptional repression domain (CRD1) by SIRT1, preferentially at Lys-1020.
Citrullinated at Arg-2142 by PADI4, which impairs methylation by CARM1 and promotes interaction with NCOA2/GRIP1.
Methylated at Arg-580 and Arg-604 in the KIX domain by CARM1, which blocks association with CREB, inhibits CREB signaling and activates apoptotic response. Also methylated at Arg-2142 by CARM1, which impairs interaction with NCOA2/GRIP1.
Sumoylated; sumoylation in the transcriptional repression domain (CRD1) mediates transcriptional repression. Desumoylated by SENP3 through the removal of SUMO2 and SUMO3.
Probable target of ubiquitination by FBXO3, leading to rapid proteasome-dependent degradation.
Phosphorylated by HIPK2 in a RUNX1-dependent manner. This phosphorylation that activates EP300 happens when RUNX1 is associated with DNA and CBFB. Phosphorylated by ROCK2 and this enhances its activity. Phosphorylation at Ser-89 by AMPK reduces interaction with nuclear receptors, such as PPARG.
Cellular localizationCytoplasm. Nucleus. In the presence of ALX1 relocalizes from the cytoplasm to the nucleus. Colocalizes with ROCK2 in the nucleus.
- Information by UniProt
- E1A-associated protein p300