2ILM

Factor Inhibiting HIF-1 Alpha D201A Mutant in Complex with FE(II), Alpha-Ketoglutarate and HIF-1 Alpha 35mer

Summary for 2ILM

• Entry DOI: 10.2210/pdb2ilm/pdb
• Related: 1H2K 1H2L 1H2M 1H2N 1YCI
• Descriptor: Hypoxia-inducible factor 1 alpha inhibitor, Hypoxia-inducible factor 1 alpha, FE (II) ION, ... (8 entities in total)
• Functional Keywords: fih, hif, dsbh, oxygenase, transcription, hypoxia, inhibitor 2-oxoglutarate, asparaginyl hydroxylase, transcription regulator, oxidoreductase
• Biological source: Homo sapiens (human); More
• Cellular location: Nucleus (Potential): Q9NWT6; Cytoplasm: Q16665
• Total number of polymer chains: 2
• Total formula weight: 45522.58

Authors

Mcdonough, M.A.,Schofield, C.J. (deposition date: 2006-10-03, release date: 2007-08-14, Last modification date: 2023-08-30)

Primary citation

Hewitson, K.S.,Holmes, S.L.,Ehrismann, D.,Hardy, A.P.,Chowdhury, R.,Schofield, C.J.,McDonough, M.A.
Evidence that two enzyme-derived histidine ligands are sufficient for iron binding and catalysis by factor inhibiting HIF (FIH).
J.Biol.Chem., 283:25971-25978, 2008

PubMed Abstract: A 2-His-1-carboxylate triad of iron binding residues is present in many non-heme iron oxygenases including the Fe(II) and 2-oxoglutarate (2OG)-dependent dioxygenases. Three variants (D201A, D201E, and D201G) of the iron binding Asp-201 residue of an asparaginyl hydroxylase, factor inhibiting HIF (FIH), were made and analyzed. FIH-D201A and FIH-D201E did not catalyze asparaginyl hydroxylation, but in the presence of a reducing agent, they displayed enhanced 2OG turnover when compared with wild-type FIH. Turnover of 2OG by FIH-D201A was significantly stimulated by the addition of HIF-1alpha(786-826) peptide. Like FIH-D201A and D201E, the D201G variant enhanced 2OG turnover but rather unexpectedly catalyzed asparaginyl hydroxylation. Crystal structures of the FIH-D201A and D201G variants in complex with Fe(II)/Zn(II), 2OG, and HIF-1alpha(786-826/788-806) implied that only two FIH-based residues (His-199 and His-279) are required for metal binding. The results indicate that variation of 2OG-dependent dioxygenase iron-ligating residues as a means of functional assignment should be treated with caution. The results are of mechanistic interest in the light of recent biochemical and structural analyses of non-heme iron and 2OG-dependent halogenases that are similar to the FIH-D201A/G variants in that they use only two His-residues to ligate iron.

PubMed: 18611856
DOI: 10.1074/jbc.M804999200

Experimental method

X-RAY DIFFRACTION (2.3 Å)