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PRC2 EZH2 (full length), (Tyr641Cys-Mutant) protein (DYKDDDDK Tag)

Origin: Human Host: Baculovirus Recombinant EAA, ScA
Catalog No. ABIN2669654
  • Target
    PRC2 EZH2
    Protein Type
    Recombinant
    Protein Characteristics
    Tyr641Cys-Mutant, full length
    Origin
    Human
    Source
    • 3
    Baculovirus
    Purification tag / Conjugate
    DYKDDDDK Tag
    Application
    Enzyme Activity Assay (EAA), Screening Assay (ScA)
    Characteristics
    Recombinant PRC2 EZH2(Y641C) Complex that includes full length EZH2 with a Y641C mutation complexed with full length SUZ12, EED and RbAp46/48 (accession numbers NP_001190176.1, NP_056170, NP_003788.2, NP_002884.1, and NP_005601.1, respectively) was expressed in Sf9 and contains an N-terminal FLAG tag at the N-terminus of EZH2. The molecular weights of expressed EZH2(Y641C), SUZ12, EED and RbAp46/48 are 87 kDa, 83 kDa, 50.2 kDa, 47.8 kDa and 47.7 kDa, respectively.
  • Application Notes
    PRC2 EZH2(Y641C) Complex is suitable for use in the study of enzyme kinetics, inhibitor screening and selectivity profiling. Specific Activity: H3K27me3 and H3K27me2 methyltransferase. HMT Assay Conditions: 50 mM TrisCl, pH 8.6, 0.02 % Triton X-100, 2 mM MgCl2, 1 mM TCEP, 100 μM SAM, 30 μM H3K27me2 peptide, 40 nM Recombinant PRC2 EZH2 (Y641C) protein at 2 hours at room temperature. Activity was detected by MALDI-TOF.
    Restrictions
    For Research Use only
  • Handling Advice
    Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
    Storage
    -80 °C
    Storage Comment
    Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation.
  • Target
    PRC2 EZH2
    Background
    PRC2 (Polycomb Repressive Complex 2) is one of the two classes Polycomb-group, or PcG proteins (the other being PRC1) that are important epigenetic determinants of stem cell identity. They play an important role in long-term epigenetic silencing of genes during cell fate determination and differentiation. PRC2 functions as a repressor of chromatin. PRC2 is required to target recruitment to specific DNA sequences (termed Polycomb Response Elements or PREs) of genomic regions to be silenced. Once associated with chromatin, the PRC2 subunit EZH2 has histone methyltransferase activity that catalyzes the trimethylation of histone H3 at Lys27. H3K27me3 is well established as a hallmark of regions of repressed chromatin. Trimethylation of Lys27 leads to the recruitment of PRC1 through the binding of H3K27me3 by chromodomain-containing proteins in PRC1. PRC1 is responsible for long-term gene silencing after cellular differentiation. Recombinant PRC2 EZH2(Y641C) Complex is a mutant version of our wild-type PRC2 Complex that contains a Tyr-to-Cys mutation at tyrosine 641 of the SET domain of EZH2. Somatic mutations of tyrosine 641, including Y641F, Y641N, Y641C, Y641S and Y641H, have been identified in patients with non-Hodgkin lymphoma and have been shown to alter substrate specificity and catalytic activity of EZH2 for histone H3 lysine 27 (H3K27) methylation states resulting in increased H3K27 trimethylation.
    Molecular Weight
    The molecular weights of expressed EZH2(Y641C), SUZ12, EED and RbAp46/48 are 87 kDa, 83 kDa, 50.2 kDa, 47.8 kDa and 47.7 kDa, respectively.
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