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Literature summary for 2.5.1.46 extracted from

  • Chawla, B.; Jhingran, A.; Singh, S.; Tyagi, N.; Park, M.H.; Srinivasan, N.; Roberts, S.C.; Madhubala, R.
    Identification and characterization of a novel deoxyhypusine synthase in Leishmania donovani (2010), J. Biol. Chem., 285, 453-463.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.5.1.46

Cloned(Commentary)

Cloned (Comment) Organism
DHSL20 is a DHS-like gene from chromosome 20 and DHS34, a DHS gene from chromosome 34, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, expression of His-tagged DHSL20 and DHS34 in Escherichia coli Leishmania donovani
expressed in Escherichia coli as a His-tagged fusion protein Leishmania donovani

Protein Variants

Protein Variants Comment Organism
L344K mutant enzyme is still devoid of any DHS activity, suggesting that other differences in amino acid sequences also contribute to its inactivity Leishmania donovani
additional information DHS34 gene deletion mutations via targeted gene replacement, chromosomal null mutants of DHS34 are only obtained in the presence of a DHS34-containing episome. HYGDELTAdhs34 and PHLEODELLTAdhs34 gene replacement cassettes are individually transfected into wild-type parasites, phenotypes, verview Leishmania donovani

Inhibitors

Inhibitors Comment Organism Structure
N-guanyl-1,7-diaminoheptane shows only little inhibitory effect on the leishmanial recombinant DHS34 Leishmania donovani

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00177
-
 [eIF5A-precursor]-L-lysine
-
 Leishmania donovani
0.00177
-
 [eIF5A-precursor]-lysine pH 9.2, 37°C, recombinant DHS34 Leishmania donovani
0.0072
-
 spermidine
-
 Leishmania donovani
0.0232
-
 spermidine pH 9.2, 37°C, recombinant DHS34 Leishmania donovani

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
42000
-
 predicted from cDNA Leishmania donovani
48000
-
 x * 48000, recombinant His-tagged DHSL20, SDS-PAGE Leishmania donovani
66000
-
 predicted from cDNA Leishmania donovani
73000
-
 2 * 66000, about, DHS34, sequence calculation, 2 * 73000, recombinant His-tagged DHS34, SDS-PAGE Leishmania donovani
137000
-
 gel filtration Leishmania donovani
137000
-
 recombinant DHS34, gel filtration Leishmania donovani

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
[eIF5A-precursor]-lysine + spermidine Leishmania donovani
-
 [eIF5A-precursor]-deoxyhypusine + propane-1,3-diamine
-
 ?
[eIF5A-precursor]-lysine + spermidine Leishmania donovani MHOM/IN/1983/AG83
-
 [eIF5A-precursor]-deoxyhypusine + propane-1,3-diamine
-
 ?

Organism

Organism UniProt Comment Textmining
Leishmania donovani
-
 two putative deoxyhypusine synthase sequences are identified in the Leishmania donovani genome
-
Leishmania donovani B5APK2 two putative deoxyhypusine synthase sequences are identified in the Leishmania donovani genome
-
Leishmania donovani B5APK2 DHS34; DHSL20 is a DHS-like gene from chromosome 20 and DHS34, a DHS gene from chromosome 34
-
Leishmania donovani MHOM/IN/1983/AG83 B5APK2 DHS34; DHSL20 is a DHS-like gene from chromosome 20 and DHS34, a DHS gene from chromosome 34
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged DHSL20 and DHS34 from Escherichia coli by metal affinity chromatography Leishmania donovani
using Ni-NTA chromatography Leishmania donovani

Source Tissue

Source Tissue Comment Organism Textmining
promastigote
-
 Leishmania donovani
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 the recombinant DHSL20 protein does not show any activity Leishmania donovani
0.0000096
-
 low specific activity of DHS34 could be due to the fact that one of the NAD+ binding regions in the DHS34 is present in the big long insertion when compared with human deoxyhypusine synthase Leishmania donovani

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information the DHSL20 protein lacks a critical lysine residue, and the recombinant protein shows no DHS activity in vitro, while DHS34 contains the critical lysine residue, and the recombinant DHS34 effectively catalyzes deoxyhypusine synthesis Leishmania donovani ?
-
 ?
additional information the DHSL20 protein lacks a critical lysine residue, and the recombinant protein shows no DHS activity in vitro, while DHS34 contains the critical lysine residue, and the recombinant DHS34 effectively catalyzes deoxyhypusine synthesis Leishmania donovani MHOM/IN/1983/AG83 ?
-
 ?
[eIF5A-precursor] + spermidine
-
 Leishmania donovani [eIF5A-precursor]-deoxyhypusine + propane-1,3-diamine
-
 ?
[eIF5A-precursor] + spermidine
-
 Leishmania donovani MHOM/IN/1983/AG83 [eIF5A-precursor]-deoxyhypusine + propane-1,3-diamine
-
 ?
[eIF5A-precursor]-L-lysine + spermidine
-
 Leishmania donovani [eIF5A-precursor]-deoxyhypusine + propane-1,3-diamine
-
 ?
[eIF5A-precursor]-L-lysine + spermidine
-
 Leishmania donovani MHOM/IN/1983/AG83 [eIF5A-precursor]-deoxyhypusine + propane-1,3-diamine
-
 ?
[eIF5A-precursor]-lysine + spermidine
-
 Leishmania donovani [eIF5A-precursor]-deoxyhypusine + propane-1,3-diamine
-
 ?
[eIF5A-precursor]-lysine + spermidine
-
 Leishmania donovani MHOM/IN/1983/AG83 [eIF5A-precursor]-deoxyhypusine + propane-1,3-diamine
-
 ?

Subunits

Subunits Comment Organism
? x * 48000, recombinant His-tagged DHSL20, SDS-PAGE Leishmania donovani
dimer 2 * 66000 Leishmania donovani
dimer 2 * 66000, about, DHS34, sequence calculation, 2 * 73000, recombinant His-tagged DHS34, SDS-PAGE Leishmania donovani
More structural modeling of DHS34, overview Leishmania donovani

Synonyms

Synonyms Comment Organism
deoxyhypusine synthase
-
 Leishmania donovani
DHS
-
 Leishmania donovani
DHS20
-
 Leishmania donovani
DHS34
-
 Leishmania donovani

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Leishmania donovani

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9.2
-
 assay at Leishmania donovani
9.6
-
 assay at Leishmania donovani

Cofactor

Cofactor Comment Organism Structure
NAD+
-
 Leishmania donovani
NAD+ dependent on Leishmania donovani

General Information

General Information Comment Organism
malfunction DHSL20 lacks the catalytic lysine residue at residue 344, which is indispensable for the enzymatic reaction and is present in the active center for catalysis, and contains leucine instead Leishmania donovani
malfunction gene deletion mutations are attempted via targeted gene replacement. Chromosomal null mutants of DHS34 could only be obtained in the presence of a DHS34-containing episome, indicating that DHS34 is an essential gene Leishmania donovani
additional information structural modeling of DHS34, overview Leishmania donovani
physiological function deoxyhypusine synthase, an NAD+-dependent enzyme, catalyzes the first step in the post-translational synthesis of an unusual amino acid, hypusine (N-(4-amino-2-hydroxybutyl)lysine), in the eukaryotic initiation factor 5A precursor protein, DHS34 is essential for Leishmania donovani Leishmania donovani