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General description
Research area: Cell signaling. Protein arginine methyltransferase 5 (PRMT5) and WD repeat-containing protein (WDR77) or methylosome protein- 50 (MEP-50) are part of the methylosome. PRMT5 is a type II PRMT enzyme. The gene encoding it is localized to human chromosome 14q11.2.Complex of human PRMT5 (GenBank Accession No. NM_006109), amino acid 2-end, with N-terminal FLAG®-tag, MW=73 kDa, and human MEP50 (GenBank Accession No. NM_024102), amino acid 2-end, with N-terminal His tag, MW=37.5 kDa, expressed in FreeStyle 293-F cells.
Application
Recombinant PRMT5/MEP50 has been used in the detection of arginine methylation in in vitro methyltransferase assay.
Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Biochem/physiol Actions
Methylosome protein 50 (MEP50) binds to the oncogenic protein arginine methyltransferase 5 (PRMT5) and forms a complex called methylosome. This complex is involved in mono- and symmetric dimethylation of arginine, which is necessary for various biological processes. MEP50 acts as a coactivator for androgen receptor (AR) and estrogen receptor (ER). PRMT5 also methylates histones and other proteins involved in genome organization, transcription, stem cells, primordial germ cells, differentiation, the cell cycle, and spliceosome assembly. The two proteins have been associated with cancer tumorigenesis and are marked as potential oncogenes.A boost in MEP50 levels is observed in several human cancers such asastrocytoma, ovarian cancer, lung cancer, glioblastoma, breast cancer,testicular tumor, and prostate cancer.
Physical form
Formulated in 40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 3 mM DTT, 80 μg/ml FLAG® peptide, and 20% glycerol.
Preparation Note
Thaw on ice. Upon first thaw, briefly spin tube containing enzyme to recover full content of the tube. Aliquot enzyme into single use aliquots. Store remaining undiluted enzyme in aliquots at -70°C. Note: Enzyme is very sensitive to freeze/thaw cycles.
Other Notes
One unit is defined as the amount of enzyme required to methylate 1 pmol of substrate/min at 37°C.
Legal Information
FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
FreeStyle is a trademark of Invitrogen Corp.
| biological source | human |
| recombinant | expressed in FreeStyle™ 293-F cells |
| assay | ≥60% (SDS-PAGE) |
| form | aqueous solution |
| mol wt | 37.5 kDa (MEP50), 73 kDa (PRMT5) |
| packaging | pkg of 20 μg |
| manufacturer/tradename | Sigma-Aldrich |
| storage condition | avoid repeated freeze/thaw cycles |
| concentration | >0.02 mg/mL |
| technique(s) | inhibition assay: suitable |
| solubility | water: soluble |
| NCBI accession no. | NM_006109 (PRMT5),NM_024102 (MEP50) |
| UniProt accession no. | O14744 |
| application(s) | life science and biopharma |
| shipped in | dry ice |
| storage temp. | −70°C |
| Gene Information | human ... PRMT5(10419) |
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