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Literature summary for 2.7.2.11 extracted from

  • Sienkiewicz, N.; Ong, H.B.; Fairlamb, A.H.
    Characterisation of a putative glutamate 5-kinase from Leishmania donovani (2018), FEBS J., 285, 2662-2678 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.2.11

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli C41(DE3) pLysS cells Leishmania donovani

Inhibitors

Inhibitors Comment Organism Structure
3,4-dehydro-L-proline 9% residual activity at 1 mM Leishmania donovani
alpha-methyl-L-proline 82% residual activity at 1 mM Leishmania donovani
cis-4-hydroxy-L-proline 87% residual activity at 1 mM Leishmania donovani
L-4-thioproline 66% residual activity at 1 mM Leishmania donovani
L-azetidine-2-carboxylic acid 25% residual activity at 1 mM Leishmania donovani
L-prolinamide 96% residual activity at 1 mM Leishmania donovani
L-proline 4% residual activity at 1 mM Leishmania donovani
L-trans-pyrrolidine-2,4-dicarboxylic acid 84% residual activity at 1 mM Leishmania donovani
Mg2+
-
 Leishmania donovani
additional information L-pipecolinic acid, D-pipecolinic acid, N-methyl-L-proline, cis-3-hydroxy-L-proline, trans-1-acetyl-4-hydroxy-L-proline, D-proline as well as L-citrulline and D,L-ornithine at 1 mM concentration do not inhibit the enzymatic conversion of L-glutamate Leishmania donovani
N-acetyl-L-proline 93% residual activity at 1 mM Leishmania donovani
trans-4-hydroxy-L-proline 89% residual activity at 1 mM Leishmania donovani

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.6
-
 ATP at pH 7.4 and 25°C Leishmania donovani
10
-
 L-glutamate at pH 7.4 and 25°C Leishmania donovani

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ 5 mM used in assay conditions Leishmania donovani

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
105000
-
 gel filtration Leishmania donovani

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-glutamate Leishmania donovani
-
 ADP + L-glutamate 5-phosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Leishmania donovani A0A0R6Y3I5
-
-

Purification (Commentary)

Purification (Comment) Organism
GST resin column chromatography Leishmania donovani

Source Tissue

Source Tissue Comment Organism Textmining
promastigote
-
 Leishmania donovani
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + glutaric acid 29% activity compared to L-glutamate Leishmania donovani ?
-
 ?
ATP + L-glutamate
-
 Leishmania donovani ADP + L-glutamate 5-phosphate
-
 ?
ATP + L-glutamate 100% activity Leishmania donovani ADP + L-glutamate 5-phosphate
-
 ?
additional information GTP, CTP or UTP do not replace ATP as phosphate donor. The enzyme has no activity with aminovaleric acid and 2-amino-4-phosphonobutyric acid Leishmania donovani ?
-
-

Subunits

Subunits Comment Organism
homotetramer 4 * 29000, SDS-PAGE Leishmania donovani
homotetramer 4 * 29033, calculated from amino acid sequence Leishmania donovani
homotetramer 4 * 29100, MALDI-TOF mass spectrometry Leishmania donovani

Synonyms

Synonyms Comment Organism
G5K
-
 Leishmania donovani
gamma-glutamyl kinase
-
 Leishmania donovani

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
11.7
-
 ATP at pH 7.4 and 25°C Leishmania donovani
12.8
-
 L-glutamate at pH 7.4 and 25°C Leishmania donovani

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
43
-
 Mg2+ at pH 7.4 and 25°C Leishmania donovani

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.39
-
 at pH 7.4 and 25°C Leishmania donovani L-proline
0.49
-
 at pH 7.4 and 25°C Leishmania donovani 3,4-dehydro-L-proline
0.73
-
 at pH 7.4 and 25°C Leishmania donovani L-azetidine-2-carboxylic acid
1.8
-
 at pH 7.4 and 25°C Leishmania donovani L-4-thioproline

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.28
-
 L-glutamate at pH 7.4 and 25°C Leishmania donovani
19.5
-
 ATP at pH 7.4 and 25°C Leishmania donovani