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Literature summary extracted from

  • Oza, S.L.; Chen, S.; Wyllie, S.; Coward, J.K.; Fairlamb, A.H.
    ATP-dependent ligases in trypanothione biosynthesis - kinetics of catalysis and inhibition by phosphinic acid pseudopeptides (2008), FEBS J., 275, 5408-5421.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.1.8 gene gspS, DNA and amino acid sequence determination and analysis, expression as His-tagged enzyme in Escherichia coli strain BL21(DE3) Crithidia fasciculata

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.3.1.8 Phosphinate behaves as a slow-binding bisubstrate inhibitor, competitive with respect to GSH and spermidine Crithidia fasciculata
6.3.1.8 phosphinate analogue of glutathionylspermidine slow tight-binding inhibition Crithidia fasciculata
6.3.1.8 phosphonate acts as a a simple ground state analogue of glutathione Crithidia fasciculata
6.3.1.8 phosphonate analogue of glutathionylspermidine classical, linear competitive inhibition with respect to GSH Crithidia fasciculata

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.1.8 0.157
-
 spermidine pH 7.2, 25°C Crithidia fasciculata
6.3.1.8 0.215
-
 ATP pH 7.2, 25°C Crithidia fasciculata
6.3.1.8 0.609
-
 glutathione pH 7.2, 25°C Crithidia fasciculata

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.3.1.8 Mg2+
-
 Crithidia fasciculata

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.1.8 glutathione + spermidine + ATP Crithidia fasciculata glutathionylspermidine is an intermediate formed in the biosynthesis of trypanothione, an essential metabolite in defence against chemical and oxidative stress in the kinetoplastida glutathionylspermidine + ADP + phosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.3.1.8 Crithidia fasciculata P90518
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.1.8 recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) to over 98% purity Crithidia fasciculata

Reaction

EC Number Reaction Comment Organism Reaction ID
6.3.1.8 glutathione + spermidine + ATP = glutathionylspermidine + ADP + phosphate the kinetic mechanism obeys a rapid equilibrium random ter-ter model Crithidia fasciculata
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.1.8 glutathione + spermidine + ATP
-
 Crithidia fasciculata glutathionylspermidine + ADP + phosphate
-
 ?
6.3.1.8 glutathione + spermidine + ATP glutathionylspermidine is an intermediate formed in the biosynthesis of trypanothione, an essential metabolite in defence against chemical and oxidative stress in the kinetoplastida Crithidia fasciculata glutathionylspermidine + ADP + phosphate
-
 ?
6.3.1.8 additional information model of ter-reactant mechanism of GspS catalysis and postulated overview Crithidia fasciculata ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
6.3.1.8 GspS
-
 Crithidia fasciculata

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.3.1.8 25
-
 assay at Crithidia fasciculata

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.3.1.8 7.2
-
 assay at Crithidia fasciculata

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.1.8 ATP
-
 Crithidia fasciculata

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
6.3.1.8 additional information
-
 additional information inhibition kinetics and mechanism, model of ter-reactant mechanism of GspS catalysis and postulated, overview Crithidia fasciculata
6.3.1.8 0.019
-
 phosphinate analogue of glutathionylspermidine pH 7.2, 25°C Crithidia fasciculata
6.3.1.8 0.029
-
 Phosphinate pH 7.2, 25°C Crithidia fasciculata
6.3.1.8 0.156
-
 phosphonate analogue of glutathionylspermidine pH 7.2, 25°C Crithidia fasciculata

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
6.3.1.8 0.072
-
 pH 7.2, 25°C Crithidia fasciculata Phosphinate