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Encompassing Amino Acids: 2-end (all components)
Applications: Useful for Western blot analysis and to investigate protein-protein interactions in the EZH2 complex.
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.
Description: 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.
Format: Aqueous buffer solution
Formulation: 25 mM Tris-HCl, pH 8.0, 138 mM NaCl, 0.05% Tween-20, and 10% glycerol.
Genbank: NM_004456 (EZH2), NM_003797(EED), NM_015355 (SUZ12), NM_153207(AEBP2), NM_005610(RbAP48)
Host Cell Line: Sf9 cells
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal His-tag
Uniprot: Q15910
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 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.
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