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Literature summary extracted from

  • Oza, S.L.; Ariyanayagam, M.R.; Fairlamb, A.H.
    Characterization of recombinant glutathionylspermidine synthetase/amidase from Crithidia fasciculata (2002), Biochem. J., 364, 679-686.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.1.78 expression in Escherichia coli, His6-tag Crithidia fasciculata
6.3.1.8 expression of a recombinant histidine-tagged enzyme in Escherichia coli Crithidia fasciculata

Protein Variants

EC Number Protein Variants Comment Organism
3.5.1.78 C79A complete loss of amidase activity without affecting synthetase activity Crithidia fasciculata
6.3.1.8 C79A mutant retains synthetase activity, but loses amidase activity Crithidia fasciculata

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.78 0.059
-
 spermidine pH 7.3, synthetase activity Crithidia fasciculata
3.5.1.78 0.114
-
 ATP pH 7.3, synthetase activity Crithidia fasciculata
3.5.1.78 0.242
-
 reduced glutathione pH 7.3, synthetase activity Crithidia fasciculata
3.5.1.78 0.5
-
 glutathionylspermidine pH 7.3, amidase activity Crithidia fasciculata
6.3.1.8 0.059
-
 spermidine pH 7.3, 25°C, synthetase activity Crithidia fasciculata
6.3.1.8 0.078
-
 spermidine pH 7.3, 25°C, synthetase activity of the C79A mutant Crithidia fasciculata
6.3.1.8 0.096
-
 MgADP- pH 7.3, 25°C, synthetase activity of the C79A mutant Crithidia fasciculata
6.3.1.8 0.114
-
 MgADP- pH 7.3, 25°C, synthetase activity Crithidia fasciculata
6.3.1.8 0.171
-
 glutathione pH 7.3, 25°C, synthetase activity of the C79A mutant Crithidia fasciculata
6.3.1.8 0.242
-
 glutathione pH 7.3, 25°C, synthetase activity Crithidia fasciculata
6.3.1.8 0.5
-
 glutathionylspermidine pH 7.3, 25°C, amidase activity Crithidia fasciculata

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.3.1.8 80520
-
 MALDI-TOF MS Crithidia fasciculata
6.3.1.8 85200
-
 gel filtration Crithidia fasciculata
6.3.1.8 86000
-
 1 * 86000, SDS-PAGE Crithidia fasciculata

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.78 Crithidia fasciculata
-
 bifunctional glutathionylspermidine synthetase, EC 6.3.1.8, and amidase, EC 3.5.1.78
-
6.3.1.8 Crithidia fasciculata P90518
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.1.78 recombinant enzyme Crithidia fasciculata
6.3.1.8 using Ni-chelating and anion-exchange chromatography Crithidia fasciculata

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.3.1.8 7.9
-
 pH 7.3, 25°C, synthetase activity Crithidia fasciculata

Storage Stability

EC Number Storage Stability Organism
6.3.1.8 -80°C, 4 months, loses less than 10% synthetase activity Crithidia fasciculata

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.78 glutathionylspermidine + H2O
-
 Crithidia fasciculata glutathione + spermidine
-
 r
3.5.1.78 trypanothione + H2O hydrolysis at 1.5% the rate of glutathionylspermidine Crithidia fasciculata glutathione + glutathionylspermidine
-
 ?
6.3.1.8 gamma-L-Glu-L-Cys-Gly + spermidine + MgATP2-
-
 Crithidia fasciculata N1-(gamma-L-Glu-L-Cys-Gly)-spermidine + MgADP- + phosphate
-
 ?
6.3.1.8 glutathionylspermidine + H2O amidase activity Crithidia fasciculata glutathione + spermidine
-
 ?

Subunits

EC Number Subunits Comment Organism
6.3.1.8 monomer 1 * 86000, SDS-PAGE Crithidia fasciculata

Synonyms

EC Number Synonyms Comment Organism
6.3.1.8 GspS
-
 Crithidia fasciculata

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.3.1.8 7.31
-
-
 Crithidia fasciculata

pH Range

EC Number pH Minimum pH Maximum Comment Organism
6.3.1.8 6.8 8
-
 Crithidia fasciculata

pI Value

EC Number Organism Comment pI Value Maximum pI Value
6.3.1.8 Crithidia fasciculata isoelectric focusing 4.7 4.6