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  • PRMT1 Methyltransferase

    Description

    PRMT1 is a major protein arginine methyltransferase (1). It specifically methylates arginine 3 (Arg 3) of H4 in vitro and in vivo. Furthermore, methylation of histone H4 at Arg 3 facilitates transcriptional activation by nuclear hormone receptor (2). Ordered cooperative functions of PRMT1, p300 and CARM1 in transcriptional activation by p53 is observed on the GADD45 gene following ectopic p53 expression and/or UV irradiation (3).

    Product Source

    PRMT1 enzyme is expressed from human PRMT1 cDNA using an MBP fusion tag expressed in E. coli.

    Properties and Usage

    Unit Definition

    One unit is defined as the amount of enzyme required to catalyze the transfer of 1 pmol of methyl group to histone H4 in a total reaction volume of 25 μl in 10 minutes at 37°C.

    Reaction Conditions

    1X HMT Reaction Buffer
    Supplement with 160 μM S-adenosylmethionine (SAM)
    Incubate at 37°C

    1X HMT Reaction Buffer:
    50 mM Tris-HCl
    5 mM MgCl2
    4 mM DTT
    pH 9 @ 25°C

    Usage Concentration

    1%

    Storage Temperature

    -20°C

    Storage Conditions

    50 mM Tris-HCl
    100 mM NaCl
    5 mM MgCl2
    4 mM DTT
    50% Glycerol
    pH 8.0 @ 25°C

    Quality Control

    Quality Assurance Statement

    • Purified free of contaminating proteases.

    Notes

    1. Storage Note: S-adenosylmethionine (SAM) is stored at -20°C as a 32 mM solution dissolved in 0.005 M sulfuric acid and 10% ethanol (pH 7.5). Under these conditions, SAM is stable for up to 6 months. SAM is unstable at 37°C and should be replenished in reactions incubated longer than 4 hours. Methylation can be optimized by using fresh SAM. Avoid freeze/thaw of enzyme.

    References

    1. Tang, J. et al. (2000). J. Biol. Chem.. 275, 7723-7730.
    2. Wang, H. et al. (2001). Science. 293, 853-857.
    3. An, W. et al. (2004). Cell. 117, 735-748.

    Supporting Documents

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.

    Datacards

    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.
    1. Can the histones be used as substrates for protein modification enzymes? Which ones?