PDBsum entry 2cgo

Oxidoreductase

PDB id: 2cgo

Contents

Protein chain

339 a.a. *

Ligands

FUM

SO4 ×4

Metals

_FE

Waters ×121

* Residue conservation analysis

PDB id:

2cgo

Name:

Oxidoreductase

Title:

Factor inhibiting hif-1 alpha with fumarate

Structure:

Hypoxia-inducible factor 1 alpha inhibitor. Chain: a. Synonym: factor inhibiting hif1, hypoxia-inducible factor asparagine hydroxylase, fih-1. Engineered: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Expressed in: escherichia coli. Expression_system_taxid: 469008.

Resolution:

2.30Å R-factor: 0.192 R-free: 0.237

Authors:

M.A.Mcdonough,I.J.Clifton,C.J.Schofield

Key ref:

K.S.Hewitson et al. (2007). Structural and mechanistic studies on the inhibition of the hypoxia-inducible transcription factor hydroxylases by tricarboxylic acid cycle intermediates. J Biol Chem, 282, 3293-3301. PubMed id: 17135241 DOI: 10.1074/jbc.M608337200

Date:

09-Mar-06 Release date: 13-Dec-06

Protein chain

Q9NWT6  (HIF1N_HUMAN) -  Hypoxia-inducible factor 1-alpha inhibitor from Homo sapiens

Seq: 349 a.a.

Struc: 339 a.a.

Enzyme reactions

• Enzyme class 1: E.C.1.14.11.30 - hypoxia-inducible factor-asparagine dioxygenase.
• Reaction: L-asparaginyl-[hypoxia-inducible factor alpha subunit] + 2-oxoglutarate + O2 = (3S)-3-hydroxy-L-asparaginyl-[hypoxia-inducible factor alpha subunit] + succinate + CO2

L-asparaginyl-[hypoxia-inducible factor alpha subunit] + 2-oxoglutarate + O2 = (3S)-3-hydroxy-L-asparaginyl-[hypoxia-inducible factor alpha subunit] + succinate + CO2 (Bound ligand (Het Group name = FUM) corresponds exactly)

• Cofactor: Fe(2+); L-ascorbate
• Enzyme class 2: E.C.1.14.11.n4 - ?????

Note, where more than one E.C. class is given (as above), each may correspond to a different protein domain or, in the case of polyprotein precursors, to a different mature protein.

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

DOI no: 10.1074/jbc.M608337200

J Biol Chem 282:3293-3301 (2007)

PubMed id: 17135241

Structural and mechanistic studies on the inhibition of the hypoxia-inducible transcription factor hydroxylases by tricarboxylic acid cycle intermediates.

K.S.Hewitson, B.M.Liénard, M.A.McDonough, I.J.Clifton, D.Butler, A.S.Soares, N.J.Oldham, L.A.McNeill, C.J.Schofield.

ABSTRACT

In humans both the levels and activity of the alpha-subunit of the hypoxia-inducible transcription factor (HIF-alpha) 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.

Selected figure(s)

Figure 1.
FIGURE 1. The reactions catalyzed by the HIF hydroxylases (PHDs and FIH).

Figure 4.
FIGURE 4. Inhibition of t-PHD2 by the TCAIs (at 1 mM final concentration). No inc refers to the assay without a preincubation step; 1hrinc refers to a 1-h preincubation of t-PHD2 with iron, buffer, and compound (where applicable), and 1hrinc + asc refers to a 1-h preincubation of t-PHD2 with iron, buffer, and ascorbate.

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2007, 282, 3293-3301) copyright 2007.

Figures were selected by an automated process.