Cloned (Comment) | Organism |
---|---|
expression of GST-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
E208A | severe impairment in binding of substrate translation initiation factor 5A, complete loss of activity | Homo sapiens |
E208A | site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding | Homo sapiens |
E208D | impairment in binding of substrate translation initiation factor 5A, retains some activity | Homo sapiens |
E208D | site-directed mutagenesis, the mutant shows reduced but not fully abolished eIF5A(deoxyhypusine) binding | Homo sapiens |
E208N | site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding | Homo sapiens |
E208Q | site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding | Homo sapiens |
E241A | severe impairment in binding of substrate translation initiation factor 5A | Homo sapiens |
E241A | site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding | Homo sapiens |
E57A | severe impairment in binding of substrate translation initiation factor 5A, complete loss of activity | Homo sapiens |
E57A | site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding | Homo sapiens |
E57D | impairment in binding of substrate translation initiation factor 5A, retains some activity | Homo sapiens |
E57D | site-directed mutagenesis, the mutant shows reduced but not fully abolished eIF5A(deoxyhypusine) binding | Homo sapiens |
E57N | site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding | Homo sapiens |
E57Q | site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) binding | Homo sapiens |
E90A | severe impairment in binding of substrate translation initiation factor 5A | Homo sapiens |
E90A | site-directed mutagenesis, the mutant shows severely impairment in eIF5A(deoxyhypusine) and iron binding | Homo sapiens |
E93A | site-directed mutagenesis, the mutant shows reduced eIF5A(deoxyhypusine) binding | Homo sapiens |
G214A | severe impairment in binding of substrate translation initiation factor 5A | Homo sapiens |
G214A | site-directed mutagenesis, the mutant shows reduced eIF5A(deoxyhypusine) but unaltered iron binding | Homo sapiens |
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 |
G63A | severe impairment in binding of substrate translation initiation factor 5A | Homo sapiens |
G63A | site-directed mutagenesis, the mutant shows reduced eIF5A(deoxyhypusine), but unaltered iron binding | Homo sapiens |
H207A | site-directed mutagenesis, the mutant is defective in iron binding | Homo sapiens |
H240A | site-directed mutagenesis, the mutant is defective in iron binding | Homo sapiens |
H56A | site-directed mutagenesis, the mutant is defective in iron binding | Homo sapiens |
H89A | site-directed mutagenesis, the mutant is defective in iron binding | Homo sapiens |
M237A | site-directed mutagenesis, the mutant shows reduced, but existing eIF5A(deoxyhypusine) and iron binding, but is catalytically inactive | Homo sapiens |
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 |
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 |
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 |
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 |
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 | Comment | Organism | Structure |
---|---|---|---|
iron chelators | - |
Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000065 | - |
6-N-[(R)-4-amino-2-hydroxybutyl]-L-lysine of human translation initiation factor 5A | pH 7.5, 37°C | Homo sapiens | |
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 | Comment | Organism | Structure |
---|---|---|---|
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 | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
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 | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Purification (Comment) | Organism |
---|---|
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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
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 | - |
? | |
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 | - |
? | |
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 | - |
? | |
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 | - |
? | |
additional information | substrate specifc bindign of recombinant wild-type and mutant enzymes, overview | Homo sapiens | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
deoxyhypusine hydroxylase | - |
Homo sapiens |
DOHH | - |
Homo sapiens |
More | the enzyme belongs to the family of 2-oxoacid- and Fe(II)-dependent dioxygenases | Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
deoxyhypusine hydroxylase assay at | Homo sapiens |
8.5 | - |
combined deoxyhypusine synthase/deoxyhypusine hydroxylase assay at | Homo sapiens |