Literature summary for 1.14.99.29 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.

Search for more literature for 1.14.99.29

Cloned(Commentary)

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

Protein Variants

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

Inhibitors	Comment	Organism	Structure
iron chelators	-	Homo sapiens

KM Value [mM]

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

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/ Products (Substrates)

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

Organism	UniProt	Comment	Textmining
Homo sapiens	-	-	-

Purification (Commentary)

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 and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
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

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

pH Optimum

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