Recombinant PHD2 (EGLN1) protein

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

Prolyl Hydroxylase Domain-Containing Protein 2 (PHD2), also known as Egl-9 Family Hypoxia-Inducible Factor 1 (EGLN1), HIF-PH2, or HPH-2, is a member of the prolyl hydroxylase domain (PHD) family of 2-oxoglutarate-dependent dioxygenases and serves as the primary cellular oxygen sensor. PHD2 catalyzes the post-translational hydroxylation of specific proline residues within the oxygen-dependent degradation (ODD) domains of hypoxia-inducible factor (HIF)-α proteins under normoxic conditions. It hydroxylates proline residues in both the N-terminal (NODD) and C-terminal (CODD) ODD domains of HIF-1α and HIF-2α, with a preference for the NODD site, particularly in HIF-1α. Hydroxylated HIF-α proteins are recognized by the von Hippel–Lindau (VHL) E3 ubiquitin ligase complex, resulting in ubiquitination and subsequent proteasomal degradation. Under hypoxic conditions, PHD2 activity is inhibited due to limited oxygen availability, allowing HIF-α proteins to escape degradation, accumulate in the cytoplasm, translocate to the nucleus, heterodimerize with HIF-1β (ARNT), and activate the transcription of genes involved in cellular adaptation to hypoxia. PHD2 is the predominant HIF prolyl hydroxylase under physiological oxygen conditions and plays a central role in maintaining oxygen homeostasis. Through regulation of HIF signaling, PHD2 controls numerous biological processes, including erythropoiesis, angiogenesis, vascular remodeling, energy metabolism, and cardiovascular development and function. Germline mutations in EGLN1 have been associated with familial erythrocytosis, underscoring the critical role of PHD2 in oxygen sensing and erythropoietin regulation. In addition to HIF-α proteins, PHD2 has been reported to hydroxylate several non-HIF substrates, although the physiological significance of many of these interactions remains under investigation. Similar to other PHD family members, PHD2 preferentially recognizes substrates containing the conserved LXXLAP motif.

Application 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 PHD2 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. 

Protein Details

Recombinant human PHD2 (EGLN1) protein was expressed in a baculovirus expression system as the full length protein (accession number NP_071334.1) with an N-terminal FLAG tag. The molecular weight of the protein is 47 kDa. 

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