Selective Inhibition of Factor Inhibiting Hypoxia-Inducible Factor

McDonough, Michael A.; McNeill, Luke A.; Tilliet, Mélanie; Papamicaël, Cyril; Chen, Qiuyun; Banerji, Biswadip; Hewitson, Kirsty S.; Schofield, Christopher J.

doi:10.1021/ja050841b

Cited by 135 publications

(183 citation statements)

References 19 publications

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“…3, panel 1) indicating that these inhibitors do not compete with CODD. A quaternary complex was also observed for N-oxalylglycine, a nonreactive 2OG analogue inhibitor of the HIF hydroxylases (44).…”

Section: Binding Of the Tcais To T-phd2 Using Electrospray Ionizationmentioning

confidence: 83%

Structural and Mechanistic Studies on the Inhibition of the Hypoxia-inducible Transcription Factor Hydroxylases by Tricarboxylic Acid Cycle Intermediates

Hewitson

Liénard

McDonough

et al. 2007

Journal of Biological Chemistry

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205

184

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In humans both the levels and activity of the ␣-subunit of the hypoxia-inducible transcription factor (HIF-␣) are regulated by its post-translation hydroxylation as catalyzed by iron-and 2-oxoglutarate (2OG)-dependent prolyl and asparaginyl hydroxylases (PHD1-3 and factor-inhibiting HIF (FIH), respectively). One consequence of hypoxia is the accumulation of tricarboxylic acid cycle intermediates (TCAIs). In vitro assays were used to assess non-2OG TCAIs as inhibitors of purified PHD2 and FIH. Under the assay conditions, no significant FIH inhibition was observed by the TCAIs or pyruvate, but fumarate, succinate, and isocitrate inhibited PHD2. Mass spectrometric analyses under nondenaturing conditions were used to investigate the binding of TCAIs to PHD2 and supported the solution studies. X-ray crystal structures of FIH in complex with Fe(II) and fumarate or succinate revealed similar binding modes for each in the 2OG co-substrate binding site. The in vitro results suggest that the cellular inhibition of PHD2, but probably not FIH, by fumarate and succinate may play a role in the Warburg effect providing that appropriate relative concentrations of the components are achieved under physiological conditions.In humans and other mammals, the ␣␤ heterodimeric hypoxia-inducible transcription factor (HIF) 4 (1) regulates the oxygen-dependent transcription of an array of genes, including those encoding for enzymes involved in glycolysis and those proteins involved in oxygen delivery. The HIF-␤ subunit (HIF-␤) is identical to aryl hydrocarbon nuclear translocator, and its levels are oxygen-independent. Both the levels and transcriptional activity of the ␣-subunit (HIF-␣) are oxygen-regulated, and hydroxylation of human HIF-␣ at one of two prolyl residues (Pro-402 or Pro-564 in human HIF-1␣) (2, 3) within the oxygen-dependent degradation domain enables binding of HIF-␣ to the von Hippel-Lindau protein, the targeting element of an ubiquitin-protein isopeptide ligase complex. Subsequent ubiquitylation leads to proteasomal degradation of HIF-␣ (for recent reviews see Refs. 4 -7). In humans this mechanism is augmented by hydroxylation of an asparaginyl residue in the C-terminal transcriptional activation domain of HIF-1␣ (8); this modification blocks the interaction of HIF-1␣ with the transcriptional co-activator CBP/p300 so disabling HIF-mediated transcription. During hypoxia, the reduction or ablation of HIF hydroxylase activity leads to accumulation of HIF-1␣, dimerization with HIF-1␤, and consequent transcription of genes involved in the hypoxic response. Hydroxylation of human HIF-1␣ is catalyzed by four Fe(II)-and 2-oxoglutarate (2OG)-dependent dioxygenases (9, 10), which use molecular oxygen and the tricarboxylic acid cycle intermediate (TCAI) 2OG as co-substrates to produce CO 2 and the TCAI succinate as co-products (Fig. 1). Of the HIF hydroxylases, i.e. three prolyl hydroxylases (PHD1, -2, and -3) (11-13) and one asparaginyl hydroxylase (factor-inhibiting HIF (FIH)) (8, 14), PHD2 is proposed to be the most important...

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Section: Binding Of the Tcais To T-phd2 Using Electrospray Ionizationmentioning

confidence: 83%

Structural and Mechanistic Studies on the Inhibition of the Hypoxia-inducible Transcription Factor Hydroxylases by Tricarboxylic Acid Cycle Intermediates

Hewitson

Liénard

McDonough

et al. 2007

Journal of Biological Chemistry

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205

184

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“…Although many 2OG oxygenase inhibitors are aromatic heterocycles (39), the only previously reported structures for 2OG oxygenases in complex with inhibitors have used N-oxalylamino acids (40), which bind iron in a bidentate manner (29,41). A series of 2-hydroxybenzoate inhibitors related to compound A have been proposed to bind to the PHDs via the carboxylate and phenolic oxygens (42), and compound A has the potential to bind in a similar bidentate manner via its amide carbonyl and phenolic oxygens.…”

Section: Resultsmentioning

confidence: 99%

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

McDonough

Flashman

et al. 2006

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

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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...

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“…Indeed, there are currently 60 known 2-oxoglutarate-dependent dioxygenases including the jumanji family of histone demethylases and the TET family of DNA-modifying proteins. Current studies are underway to determine the contribution of these proteins to the salubrious effects of global PHD inhibitors (Mole et al, 2003;Mecinovic et al, 2009;Rose et al, 2010Rose et al, , 2011McDonough et al, 2005). The most compelling data to date that HIF PHDs can directly participate in cell death induced by injury stimuli found in stroke has come from in vitro studies.…”

Section: Hypoxia-inducible Factor Prolyl Hydroxylase Inhibition As Anmentioning

confidence: 99%

Hypoxia-Inducible Factor Prolyl Hydroxylase Inhibition: Robust New Target or Another Big Bust for Stroke Therapeutics?

Karuppagounder

Ratan

2012

J Cereb Blood Flow Metab

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A major challenge in developing stroke therapeutics that augment adaptive pathways to stress has been to identify targets that can activate compensatory programs without inducing or adding to the stress of injury. In this regard, hypoxia-inducible factor prolyl hydroxylases (HIF PHDs) are central gatekeepers of posttranscriptional and transcriptional adaptation to hypoxia, oxidative stress, and excitotoxicity. Indeed, some of the known salutary effects of putative 'antioxidant' iron chelators in ischemic and hemorrhagic stroke may derive from their abilities to inhibit this family of iron, 2-oxoglutarate, and oxygen-dependent enzymes. Evidence from a number of laboratories supports the notion that HIF PHD inhibition can improve histological and functional outcomes in ischemic and hemorrhagic stroke models. In this review, we discuss this evidence and highlight important gaps in our understanding that render HIF PHD inhibition a promising but not yet preclinically validated target for protection and repair after stroke.

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Selective Inhibition of Factor Inhibiting Hypoxia-Inducible Factor

Cited by 135 publications

References 19 publications

Structural and Mechanistic Studies on the Inhibition of the Hypoxia-inducible Transcription Factor Hydroxylases by Tricarboxylic Acid Cycle Intermediates

Structural and Mechanistic Studies on the Inhibition of the Hypoxia-inducible Transcription Factor Hydroxylases by Tricarboxylic Acid Cycle Intermediates

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

Hypoxia-Inducible Factor Prolyl Hydroxylase Inhibition: Robust New Target or Another Big Bust for Stroke Therapeutics?

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