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Recombinant Proteins & Enzymes

Recombinant PHD3 (EGLN3) protein

Catalog No: 81033 Format: 50 µg $395 Buy Now
Catalog No: 81733 Format: 1 mg $2,600 Buy Now
Expressed In: E. coli Protein Species: Human

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for Recombinant PHD3 (EGLN3) proteinContents

Recombinant PHD3 (EGLN3) protein is supplied in 25 mM Tris-HCl pH 8.0, 300 mM NaCl, 10% glycerol and 0.5 mM TCEP. Please refer to product insert upon arrival for lot-specific concentration.

for Recombinant PHD3 (EGLN3) proteinBackground

Prolyl Hydroxylase Domain-Containing Protein 3 (PHD3) or Egl-9 Family Hypoxia Inducible Factor 3 (EGLN3), also known as PHIF-PH3 or HPH-3, is a prolyl hydroxylase. PHD3 is a cellular oxygen sensor that catalyzes the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins under normoxic conditions. It can hydroxylate a specific proline found in each of the oxygen-dependent degradation (ODD) domains (N-terminal, NODD, and C-terminal, CODD) of HIF1A, also HIF2A. PHD3 has a preference for the CODD site for both HIF1A and HIF2A. Hydroxylation on the NODD site by PHD3 appears to require prior hydroxylation on the CODD site, and then hydroxylated HIFs are targeted for proteasomal degradation via the von Hippel-Lindau ubiquitination complex. Under hypoxic conditions, the hydroxylation reaction is attenuated allowing HIFs to escape degradation resulting in their translocation to the nucleus, heterodimerization with HIF1B, and increased expression of hypoxy-inducible genes. PHD3 can also hydroxylate PKM in hypoxia, limiting glycolysis, while under normoxia, hydroxylate and regulates the stability of ADRB2. In cardiomyocytes, it inhibits the anti-apoptotic effect of BCL2 by disrupting the BAX-BCL2 complex. In neurons, it has a NGF-induced proapoptotic effect, probably through regulating CASP3 activity. PHD3 is essential for hypoxic regulation of neutrophilic inflammation. Beside, it plays a crucial role in DNA damage response (DDR) by hydroxylating TELO2, promoting its interaction with ATR which is required for activation of the ATR/CHK1/p53 pathway. Its target proteins are preferentially recognized via a LXXLAP motif. 

for Recombinant PHD3 (EGLN3) proteinApplication Notes

This protein is suitable for use in binding assays, inhibitor screening, and selectivity profiling.

Assay Conditions: 3 µM HIF1A (HIF-1α) peptide (DLDLEALAPYIPADDDFQL) was incubated with 300 nM PHD3 protein in 30 µl reaction system containing 20 mM Tris-HCl pH 7.5, 5 mM KCl, 1.5 mM MgCl2, 1 mM DTT, 100 µM 2-oxoglutarate, 100 µM ascorbate and 50 µM (NH4)2Fe(SO4)2·6H2O for 2 hours at 30°C. MALDI-TOF was used for detection. 

for Recombinant PHD3 (EGLN3) proteinProtein Details

Recombinant human PHD3 (EGLN3) protein was expressed in E.coli as the full length protein (accession number NP_071356.1) with an N-terminal 6× His tag. The molecular weight of the protein is 31.1 kDa. Assay Conditions: 3 µM HIF1A (HIF-1α) peptide (DLDLEALAPYIPADDDFQL) was incubated with 300 nM PHD3 protein in 30 µl reaction system containing 20 mM Tris-HCl pH 7.5, 5 mM KCl, 1.5 mM MgCl2, 1 mM DTT, 100 µM 2-oxoglutarate, 100 µM ascorbate and 50 µM (NH4)2Fe(SO4)2·6H2O for 2 hours at 30°C. MALDI-TOF was used for detection. 

 
Recombinant PHD3 / EGLN3 protein

Recombinant PHD3 / EGLN3 protein
10% SDS-PAGE Coomassie staining.

MW: 31.1 kDa

Purity: >75% 

MALDI-TOF Assay for Recombinant PHD3 / EGLN3 protein

MALDI-TOF Assay for Recombinant PHD3 / EGLN3 protein
3 µM HIF-1 α peptide was incubated with 300 nM PHD3 protein in 30 µl reaction system containing 20 mM Tris-HCl pH 7.5, 5 mM KCl, 1.5 mM MgCl2, 1 mM DTT, 100 µM 2-oxoglutarate, 100 µM ascorbate and 50 µM (NH4)2Fe(SO4)2·6H2O for 2 hours at 30°C.

MALDI-TOF was used for detection.

for Recombinant PHD3 (EGLN3) proteinStorage

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.

for Recombinant PHD3 (EGLN3) proteinGuarantee

This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.

 

Technical Data Sheet

Recombinant PHD3 (EGLN3) protein
 

Data Thumbnails

Recombinant PHD3 / EGLN3 protein

Recombinant PHD3 / EGLN3 protein
(Click to enlarge and view details).

MALDI-TOF Assay for Recombinant PHD3 / EGLN3 protein

MALDI-TOF Assay for Recombinant PHD3 / EGLN3 protein
(Click to enlarge and view details).