Recombinant BRD3 (24-144) protein

Aliases: ORFX; RING3L
Catalog No: 31379 Format: 100 µg ¥3,200 Buy
Catalog No: 31779 Format: 1 mg ¥20,280 Buy

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Expressed In: E. coli Protein Species: Human

Contents

A representative Technical Data Sheet (TDS) is provided here. Please refer to the lot-specific TDS you will receive with your order for the lot-specific buffer contents and protein concentration.

Background

Bromodomain-containing protein 3 (BRD3), also known as RING3L, belongs to the BET subclass of proteins, which are characterized by two N-terminal bromodomains and one ET (Extra Terminal) domain. BRDs associate with chromatin through their bromodomains that recognize acetylated histone lysine residues. Bromodomains function as ‘readers’ of these epigenetic histone marks and regulate chromatin structure and gene expression by linking associated proteins to the acetylated nucleosomal targets. The ET domain functions as a protein binding motif and exerts atypical serine-kinase activity. The BET family consists of at least four members in mouse and human, BRD2 (also referred to as FSRG1, RING3), BRD3 (FSRG2, ORFX), BRD4 (FSRG4, MCAP/HUNK1), and BRDT (FSRG3, BRD6). BRD3 binds and regulates GATA1 in an acetylation-dependent manner. GATA1 is a key regulator of gene expression for erythroid and megakaryocyte-specific genes, and mutations in GATA1 have been associated with congenital anemias and megakaryoblastic leukemias. Interestingly, tight interaction of BRD3 with GATA1 requires multiple acetylation modifications, and structural data showed that two adjacent acetylation sites in GATA1 interact with a single bromodomain. BRD3 protein expression is induced in activated lymphocytes. Additionally, it is highly expressed in undifferentiated ES cells and expression is observed to drop upon endothelial differentiation. Altered expression levels of BRD3 have been observed in certain cancers, such as nasopharyngeal carcinomas and bladder cancer. BRD3 also interacts with LANA-1, the Kaposi's sarcoma-associated herpesvirus (KSHV) latency-associated nuclear antigen 1, which is required for the replication of episomal viral genomes.

Application Notes

Recombinant BRD3 (24-144) is suitable for use in binding assays, inhibitor screening, and selectivity profiling.

Protein Details

The peptide corresponding to amino acids 24 - 144 that contains the bromodomain sequences of BRD3 (accession number NM_007371.3) was expressed in E. coli and contains an N-terminal His-Tag and C-terminal FLAG-Tag with an observed molecular weight of 20.5 kDa. It shows binding specificity for acetylated H3K18, H4K12, H4K20 and H4K12/K16/K20. The recombinant protein is >90% pure by SDS-PAGE.

 

Recombinant BRD3 (24-144) protein gel.
BRD3 (24-144) protein was run on an SDS-PAGE gel and stained with Coomassie Blue.

HTRF assay for Recombinant BRD3 (24-144) activity.
3.3 µM histone peptide H4K5/8/12/16 (4Ac) was incubated with BRD3 (24-144) protein in reaction buffer including 50mM HEPES-NaOH pH 7.0, 0.1% BSA for 1 hour at room temperature. Anti-Flag antibody was used to detect reaction products.

HTRF assay for Recombinant BRD3 (24-144) activity.
3.3 µM histone peptide H4K5/8/12/16 (4Ac) was incubated with BRD3 (24-144) protein in reaction buffer including 50mM HEPES-NaOH pH 7.0, 0.1% BSA for 1 hour at room temperature. Anti-Flag antibody was used to detect reaction products.

Storage

Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.

Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.