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Literature summary extracted from

  • Kang, K.R.; Kim, Y.S.; Wolff, E.C.; Park, M.H.
    Specificity of the deoxyhypusine hydroxylase-eukaryotic translation initiation factor (eIF5A) interaction: identification of amino acid residues of the enzyme required for binding of its substrate, deoxyhypusine-containing eIF5A (2007), J. Biol. Chem., 282, 8300-8308.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.99.29 expression of GST-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
1.14.99.29 E208A severe impairment in binding of substrate translation initiation factor 5A, complete loss of activity Homo sapiens
1.14.99.29 E208A site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding Homo sapiens
1.14.99.29 E208D impairment in binding of substrate translation initiation factor 5A, retains some activity Homo sapiens
1.14.99.29 E208D site-directed mutagenesis, the mutant shows reduced but not fully abolished eIF5A(deoxyhypusine) binding Homo sapiens
1.14.99.29 E208N site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding Homo sapiens
1.14.99.29 E208Q site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding Homo sapiens
1.14.99.29 E241A severe impairment in binding of substrate translation initiation factor 5A Homo sapiens
1.14.99.29 E241A site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding Homo sapiens
1.14.99.29 E57A severe impairment in binding of substrate translation initiation factor 5A, complete loss of activity Homo sapiens
1.14.99.29 E57A site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding Homo sapiens
1.14.99.29 E57D impairment in binding of substrate translation initiation factor 5A, retains some activity Homo sapiens
1.14.99.29 E57D site-directed mutagenesis, the mutant shows reduced but not fully abolished eIF5A(deoxyhypusine) binding Homo sapiens
1.14.99.29 E57N site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding Homo sapiens
1.14.99.29 E57Q site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding Homo sapiens
1.14.99.29 E90A severe impairment in binding of substrate translation initiation factor 5A Homo sapiens
1.14.99.29 E90A site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) and iron binding Homo sapiens
1.14.99.29 E93A site-directed mutagenesis, the mutant shows reduced eIF5A(deoxyhypusine) binding Homo sapiens
1.14.99.29 G214A severe impairment in binding of substrate translation initiation factor 5A Homo sapiens
1.14.99.29 G214A site-directed mutagenesis, the mutant shows reduced eIF5A(deoxyhypusine) but unaltered iron binding Homo sapiens
1.14.99.29 G247A site-directed mutagenesis, the mutant shows 50-80% reduced activity compared to the wild-type enzyme, but is capable of binding both iron and substrate Homo sapiens
1.14.99.29 G63A severe impairment in binding of substrate translation initiation factor 5A Homo sapiens
1.14.99.29 G63A site-directed mutagenesis, the mutant shows reduced eIF5A(deoxyhypusine), but unaltered iron binding Homo sapiens
1.14.99.29 H207A site-directed mutagenesis, the mutant is defective in iron binding Homo sapiens
1.14.99.29 H240A site-directed mutagenesis, the mutant is defective in iron binding Homo sapiens
1.14.99.29 H56A site-directed mutagenesis, the mutant is defective in iron binding Homo sapiens
1.14.99.29 H89A site-directed mutagenesis, the mutant is defective in iron binding Homo sapiens
1.14.99.29 M237A site-directed mutagenesis, the mutant shows reduced, but existing eIF5A(deoxyhypusine) and iron binding, but is catalytically inactive Homo sapiens
1.14.99.29 Q215 site-directed mutagenesis, the mutant shows 50-80% reduced activity compared to the wild-type enzyme, but is capable of binding both iron and substrate Homo sapiens
1.14.99.29 R183a site-directed mutagenesis, the mutant shows 50-80% reduced activity compared to the wild-type enzyme, but is capable of binding both iron and substrate Homo sapiens
1.14.99.29 R26A site-directed mutagenesis, the mutant shows 50-80% reduced activity compared to the wild-type enzyme, but is capable of binding both iron and substrate Homo sapiens
1.14.99.29 R88A site-directed mutagenesis, the mutant shows 50-80% reduced activity compared to the wild-type enzyme, but is capable of binding both iron and substrate Homo sapiens
1.14.99.29 S202A site-directed mutagenesis, the mutant shows 50-80% reduced activity compared to the wild-type enzyme, but is capable of binding both iron and substrate Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.99.29 iron chelators
-
 Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.99.29 0.000065
-
 6-N-[(R)-4-amino-2-hydroxybutyl]-L-lysine of human translation initiation factor 5A pH 7.5, 37°C Homo sapiens
1.14.99.29 0.000376
-
 6-N-[(R)-4-amino-2-hydroxybutyl]-L-lysine of yeast translation initiation factor 5A pH 7.5, 37°C Homo sapiens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.14.99.29 Fe2+ a non-heme iron enzyme, DOHH probably contains one binuclear, diiron, active center, iron-binding active site structure of DOHH and mode of iron binding, the strictly conserved His-Glu motifs His56-Glu57, His89-Glu90, His207-Glu208 and His240-Glu241, are involved in iron binding, overview Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.99.29 eIF5A-Nepsilon-(4-aminobutyl)lysine + AH2 + O2 Homo sapiens the enzyme catalyzes the final step of the post-translational synthesis of hypusine, i.e. Nepsilon-(4-amino-2-hydroxybutyl)lysine, in the eukaryotic translation initiation factor 5A, eIF5A eIF5A-Nepsilon-(4-amino-2-hydroxybutyl)lysine + A + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.14.99.29 Homo sapiens
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.99.29 recombinant GST-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by glutathione affinity chromatography, the tag is cleaved off by thrombin Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.99.29 6-N-[(R)-4-amino-2-hydroxybutyl]-L-lysine of human translation initiation factor 5A + electron donor + O2 strong preference of enzyme for binding the 6-N-(4-amino-2-hydroxybutyl)-L-lysine-containing form of translation initiation factor 5A Homo sapiens 6-N-(4-aminobutyl)-L-lysine of human translation initiation factor 5A + oxidized electron donor + H2O
-
 ?
1.14.99.29 6-N-[(R)-4-amino-2-hydroxybutyl]-L-lysine of yeast translation initiation factor 5A + electron donor + O2 strong preference of enzyme for binding the 6-N-(4-amino-2-hydroxybutyl)-L-lysine-containing form of translation initiation factor 5A Homo sapiens 6-N-(4-aminobutyl)-L-lysine of yeast translation initiation factor 5A + oxidized electron donor + H2O
-
 ?
1.14.99.29 eIF5A-Nepsilon-(4-aminobutyl)lysine + AH2 + O2 the enzyme catalyzes the final step of the post-translational synthesis of hypusine, i.e. Nepsilon-(4-amino-2-hydroxybutyl)lysine, in the eukaryotic translation initiation factor 5A, eIF5A Homo sapiens eIF5A-Nepsilon-(4-amino-2-hydroxybutyl)lysine + A + H2O
-
 ?
1.14.99.29 eIF5A-Nepsilon-(4-aminobutyl)lysine + AH2 + O2 eIF5A is the eukaryotic translation initiation factor 5A, specificity of the interaction between eIF5A and DOHH using the isoform eIF5A-1, DOHH displays a strong preference for binding the deoxyhypusine-containing form of eIF5A, over the eIF5A precursor or the hypusine-containing eIF5A, the deoxyhypusine side chain of the substrate is primarily anchored by gamma-carboxyl groups of Glu57 and Glu208 at the DOHH active site, substrate binding modelling, overview Homo sapiens eIF5A-Nepsilon-(4-amino-2-hydroxybutyl)lysine + A + H2O
-
 ?
1.14.99.29 additional information substrate specifc bindign of recombinant wild-type and mutant enzymes, overview Homo sapiens ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.14.99.29 deoxyhypusine hydroxylase
-
 Homo sapiens
1.14.99.29 DOHH
-
 Homo sapiens
1.14.99.29 More the enzyme belongs to the family of 2-oxoacid- and Fe(II)-dependent dioxygenases Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.99.29 37
-
 assay at Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.99.29 7.5
-
 deoxyhypusine hydroxylase assay at Homo sapiens
1.14.99.29 8.5
-
 combined deoxyhypusine synthase/deoxyhypusine hydroxylase assay at Homo sapiens