The use of dioxygen by HIF prolyl hydroxylase (PHD1)

McNeill, Luke A.; Hewitson, Kirsty S.; Gleadle, Jonathan; Horsfall, Louise; Oldham, Neil J.; Maxwell, Patrick H.; Pugh, Christopher W.; Ratcliffe, Peter J.; Schofield, Christopher J.

doi:10.1016/s0960-894x(02)00219-6

Cited by 103 publications

(79 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…8, which is published as supporting information on the PNAS web site) and from mutagenesis of Arg-383 to Ala leading to almost complete inactivation of the protein (Ͻ5% by 2OG turnover assays with CODDD 556 -574 peptide substrate) (unpublished data). The assignment of Arg-383 as a 2OG-binding residue confirms a prior study in which mutation of the equivalent Arg in PHD1 (Arg-367) to Ala was shown to ablate activity (19).…”

Section: Resultssupporting

confidence: 84%

Cellular oxygen sensing: Crystal structure of hypoxia-inducible factor prolyl hydroxylase (PHD2)

McDonough

Flashman

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

332

387

View full text Add to dashboard Cite

Cellular and physiological responses to changes in dioxygen levels in metazoans are mediated via the posttranslational oxidation of hypoxia-inducible transcription factor (HIF). Hydroxylation of conserved prolyl residues in the HIF-␣ subunit, catalyzed by HIF prolyl-hydroxylases (PHDs), signals for its proteasomal degradation. The requirement of the PHDs for dioxygen links changes in dioxygen levels with the transcriptional regulation of the gene array that enables the cellular response to chronic hypoxia; the PHDs thus act as an oxygen-sensing component of the HIF system, and their inhibition mimics the hypoxic response. We describe crystal structures of the catalytic domain of human PHD2, an important prolyl-4-hydroxylase in the human hypoxic response in normal cells, in complex with Fe(II) and an inhibitor to 1.7 Å resolution. PHD2 crystallizes as a homotrimer and contains a double-stranded ␤-helix core fold common to the Fe(II) and 2-oxoglutarate-dependant dioxygenase family, the residues of which are well conserved in the three human PHD enzymes (PHD 1-3). The structure provides insights into the hypoxic response, helps to rationalize a clinically observed mutation leading to familial erythrocytosis, and will aid in the design of PHD selective inhibitors for the treatment of anemia and ischemic disease.erythropoietin ͉ dioxygenase ͉ hypoxic response ͉ 2-oxoglutarate I n metazoans the ␣͞␤ heterodimeric hypoxia-inducible transcription factor (HIF) (1) regulates the transcription of an array of genes including those coding for glycolytic enzymes, erythropoietin, and VEGF. The levels and transcriptional activity of the HIF-␣, but not the HIF-␤, subunit are regulated by oxygen. Hydroxylation of either Pro-402 or Pro-564 in human HIF-1␣ (2, 3) within the C-terminal oxygen-dependent degradation domain (CODDD) enables its binding to the von Hippel-Lindau protein (pVHL), a targeting element of the E3-ubiquitin ligase; subsequent ubiquitylation leads to proteasomal degradation of HIF-␣ (for reviews, see refs. 4 -7). In humans, this mechanism is augmented by hydroxylation of an asparagine residue in the C-terminal transcriptional activation domain (8); this modification blocks interaction of HIF-1␣ with the CBP͞p300 coactivator, thereby disabling HIFmediated transcription.Hydroxylation of HIF-1␣ is catalyzed by four 2-oxoglutarate (2OG) dioxygenases: three prolyl hydroxlyases (PHD 1, 2, and 3) (also known as HPH 3, 2, and 1 and EGLN 2, 1, and 3; refs. 9-11) and an asparaginyl hydroxylase [factor inhibiting HIF (FIH); refs. 12 and 13]. The available evidence implicates PHD2 as the most important HIF hydroxylase in down-regulating the hypoxic response during normoxia (5,7,14,15).The HIF hydroxylases are Fe(II) and 2OG-dependent dioxygenases (16, 17); their requirement for dioxygen has led to their characterization as cellular oxygen sensors (refs. 9 -11, 18, and 19; Fig. 1a). The first 2OG dioxygenase to be identified was procollagen prolyl-hydroxylase, which like the PHDs catalyzes trans-4-hydroxylation reactions. Pro...

show abstract

Section: Resultssupporting

confidence: 84%

Cellular oxygen sensing: Crystal structure of hypoxia-inducible factor prolyl hydroxylase (PHD2)

McDonough

Flashman

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

332

387

View full text Add to dashboard Cite

show abstract

“…In the cases of ANS (with (±)-naringenin as substrate) and those of PHD2 and FIH, the level of incorporation of 18 O into the enzymatic products was also analysed. In each case it was found to be >99% in accord with literature results [13][14][15]. The corresponding analysis was not carried out with AlkB as the oxygenated product, formaldehyde, undergoes facile exchange [22,23].…”

Section: Resultssupporting

confidence: 81%

Incorporation of oxygen into the succinate co‐product of iron(II) and 2‐oxoglutarate dependent oxygenases from bacteria, plants and humans

et al. 2005

View full text Add to dashboard Cite

The ferrous iron and 2-oxoglutarate (2OG) dependent oxygenases catalyse two electron oxidation reactions by coupling the oxidation of substrate to the oxidative decarboxylation of 2OG, giving succinate and carbon dioxide coproducts. The evidence available on the level of incorporation of one atom from dioxygen into succinate is inconclusive. Here, we demonstrate that five members of the 2OG oxygenase family, AlkB from Escherichia coli, anthocyanidin synthase and flavonol synthase from Arabidopsis thaliana, and prolyl hydroxylase domain enzyme 2 and factor inhibiting hypoxia-inducible factor-1 from Homo sapiens all incorporate a single oxygen atom, almost exclusively derived from dioxygen, into the succinate co-product.

show abstract

“…Then, von Hippel-Lindau protein (pVHL), a part of the E3 ubiquitin ligase protein complex, binds to the modified HIF-1·, which results in the ubiquitination and proteasomal degradation of HIF-1· (15). Since the enzymatic reaction of prolyl hydroxylation requires molecular oxygen and iron, hypoxia or iron deficiency limit this hydroxylation, thereby precluding the binding of pVHL and thus leading to the stabilization of HIF-1· (16).…”

Section: Introductionmentioning

confidence: 99%

A domain responsible for HIF-1α degradation by YC-1, a novel anticancer agent

Kim

Yeo

Chun³

et al. 2006

Int J Oncol

View full text Add to dashboard Cite

Abstract. HIF-1· is believed to promote tumor growth and metastasis, and many efforts have been made to develop new anticancer agents based on HIF-1· inhibition. YC-1 is a widely used HIF-1· inhibitor both in vitro and in vivo, and is being developed as a novel class of anticancer drug. However, little is known about the mechanism by which YC-1 degrades HIF-1·. As the first step for understanding the mechanism of action of YC-1, we here identified the HIF-1· domain responsible for YC-1-induced protein degradation. YC-1 blocked the HIF-1· induction by hypoxia, iron chelation, and proteasomal inhibition and also degraded ectopically expressed HIF-1·. In deletion analyses, C-terminal HIF-1· was found to be sensitively degraded by YC-1. Using a GFP-fusion method, the YC-1-induced degradation domain was identified as the aa. 720-780 region of HIF-1·. We next tested the possible involvement of HDAC7 or OS-9 in YC-1-induced HIF-1· degradation. However, their binding to HIF-1· was not affected by YC-1, suggesting that they are not involved in the YC-1 action. It is also suggested that YC-1 targets a novel pathway regulating HIF-1· stability.

show abstract

The use of dioxygen by HIF prolyl hydroxylase (PHD1)

Cited by 103 publications

References 27 publications

Cellular oxygen sensing: Crystal structure of hypoxia-inducible factor prolyl hydroxylase (PHD2)

Cellular oxygen sensing: Crystal structure of hypoxia-inducible factor prolyl hydroxylase (PHD2)

Incorporation of oxygen into the succinate co‐product of iron(II) and 2‐oxoglutarate dependent oxygenases from bacteria, plants and humans

A domain responsible for HIF-1α degradation by YC-1, a novel anticancer agent

Contact Info

Product

Resources

About