Mutant version of our regular EZH2 5-member complex (Cat. #51004), but with a Tyr-to-Phe mutation on a.a. 646 of the EZH2 protein. Complex of human EZH2 (GenBank Accession No. NM_004456), a.a. 2-end; Y646F*, with N-terminal His tag, MW= 86 kDa, human EED (NM_003797), a.a. 2-end with N-terminal Flag tag, MW= 51 kDa, human SUZ12 (NM_015355), a.a. 2-end with N-terminal His tag, MW = 87 kDa, Human AEBP2 (NM_153207), a.a. 2-end with N-terminal His tag, MW= 53 kDa, and human RbAp48 (NM_005610), a.a. 2-end with N-terminal His tag, MW = 48 kDa, co-expressed in baculovirus expression system.
Host cell line: Sf9 cells
Background
EZH2 is the catalytic subunit of the PRC2/EED-EZH2 complex, which methylates Lys9 (H3K9me) and Lys27 (H3K27me) of histone H3, leading to transcriptional repression of the affected target gene. Able to mono-, di- and trimethylate Lys27 of histone H3 to form H3K27me1, H3K27me2 and H3K27me3, respectively. Compared to EZH2-containing complexes, it is more abundant in embryonic stem cells and plays a major role in forming H3K27me3, which is required for embryonic stem cell identity and proper differentiation. The PRC2/EED-EZH2 complex may also serve as a recruiting platform for DNA methyltransferases, thereby linking two epigenetic repression systems. Genes repressed by the PRC2/EED-EZH2 complex include HOXC8, HOXA9, MYT1, CDKN2A and retinoic acid target genes. EZH2 can also methylate non-histone proteins such as the transcription factor GATA4.
Immunogen Region
2-end (all components)
Tag
N-terminal His-tag
Formulation
25 mM Tris-HCl, pH 8.0, 138 mM NaCl, 0.05% Tween-20, and 10% glycerol.
Species
Human
Application
Useful for Western blot analysis and to investigate protein-protein interactions in the EZH2 complex.
Notes
This mutation is often cited in the scientific literature as Y641F in variant 3 (GenBank #NM_001203247). Avoid freeze/thaw cycles.
KTM6A, EZH2, PRC2, Enhancer Of Zeste Homolog 2, EZH2 (Y646F), EZH , Retinoblastoma Binding Protein 4 (RBBP4)
Uniprot
Q15910
Shipping Temperature
-80°C (dry ice)
Format
Aqueous buffer solution
Storage
At least 6 months at -80°C.
Reference
1. Varambally, S. et al. Science 322 (5908), 1695-1699 (2008). 2. Rakotobe, D. et al., Virol. J. 5, 32 (2008).Application Reference(s): 1. Arcand, M., et al., "Development of High-Throughput Assays to Study Methylases, Demethylases and Deacetylases." Poster presented as part of the BioTechniques Fall 2011 Epigenetics Meeting. 2. Yap, D.B, et al., "Somatic mutations at EZH2 Y641 act dominantly through a mechanism of selectively altered PRC2 catalytic activity, to increase H3K27 trimethlyation." (2010). Blood. 117: 2451-2459.