Literature summary for 6.3.1.8 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.

Search for more literature for 6.3.1.8

Cloned(Commentary)

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

Inhibitors

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

KM Value [mM]

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

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	-	Crithidia fasciculata

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
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

Organism	UniProt	Comment	Textmining
Crithidia fasciculata	P90518	-	-

Purification (Commentary)

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

Reaction

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

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
glutathione + spermidine + ATP	-	Crithidia fasciculata	glutathionylspermidine + ADP + phosphate	-	?
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	-	?
additional information	model of ter-reactant mechanism of GspS catalysis and postulated overview	Crithidia fasciculata	?	-	?

Synonyms

Synonyms	Comment	Organism
GspS	-	Crithidia fasciculata

Temperature Optimum [°C]

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

pH Optimum

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

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Crithidia fasciculata

Ki Value [mM]

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

IC50 Value

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

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Release 2023.1 (January 2023)