Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Janowiak, B.E.; Hayward, M.A.; Peterson, F.C.; Volkman, B.F.; Griffith, O.W.
    gamma-Glutamylcysteine synthetase-glutathione synthetase: domain structure and identification of residues important in substrate and glutathione binding (2006), Biochemistry, 45, 10461-10473.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.2.2
-
 Streptococcus agalactiae
6.3.2.3
-
 Streptococcus agalactiae

Protein Variants

EC Number Protein Variants Comment Organism
6.3.2.2 D520stop KM-value for L-Glu is 1.3fold higher than wild-type value, KM-values for L-Cys and ATP are nearly identical to wild-type value, Ki-value for GSH is 2.2fold higher than wild-type value, Ki-value for gamma-glutamylcysteine is 1.3fold higher than wild-type value Streptococcus agalactiae
6.3.2.2 E494stop KM-value for L-Glu, L-Cys and ATP are nearly identical to wild-type value, Ki-value for GSH is 7.7fold lower than wild-type value, Ki-value for gamma-glutamylcysteine is 2.4fold higher than wild-type value Streptococcus agalactiae
6.3.2.2 G441stop KM-value for L-Glu is 3.5fold higher than wild-type value, KM-values for L-Cys and ATP are nearly identical to wild-type value Streptococcus agalactiae
6.3.2.2 H144A KM-value for L-Glu is 10fold higher than wild-type value, KM-value for L-Cys is 6.4fold higher than wild-type value, KM-value for ATP is nearly identical to wild-type value Streptococcus agalactiae
6.3.2.2 K526A KM-value for L-Glu, L-Cys and ATP are nearly identical to wild-type value Streptococcus agalactiae
6.3.2.2 additional information wild-type enzyme is nearly uninhibited by GSH, shorter gamma-glutamylcysteine synthetase domain constructs are strongly inhibited. Chimeras of Streptococcus agalactiae gamma-glutamylcysteine synthetase-glutathione synthetase are made containing gamma-glutamylcysteine synthetase domain flexible loop sequences from Enterococcus faecalis and Pasteurella multocida gamma-glutamylcysteine synthetase-glutathione synthetase isoforms that are inhibited by GSH. Inhibition remains Streptococcus agalactiae-like (very weak) Streptococcus agalactiae
6.3.2.2 R508stop KM-value for L-Glu is 1.6fold higher than wild-type value, KM-value for L-Cys is 1.7fold higher than wild-type value, KM-value for ATP is nearly identical to wild-type value, Ki-value for GSH is 2fold higher than wild-type value, Ki-value for gamma-glutamylcysteine is 3.3fold higher than wild-type value Streptococcus agalactiae
6.3.2.2 Y464stop KM-values for L-Glu, L-Cys and ATP are nearly identical to wild-type value, Ki-value for GSH is 11fold lower than wild-type value, Ki-value for gamma-glutamylcysteine is 1.3fold lower than wild-type value Streptococcus agalactiae
6.3.2.3 D448A low activity Streptococcus agalactiae
6.3.2.3 H144A higher activity than the wild type enzyme Streptococcus agalactiae
6.3.2.3 K485A very low activity Streptococcus agalactiae
6.3.2.3 K489A low activity Streptococcus agalactiae
6.3.2.3 K526A very low activity Streptococcus agalactiae

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.3.2.2 gamma-glutamylcysteine
-
 Streptococcus agalactiae
6.3.2.2 GSH wild-type enzyme is nearly uninhibited by GSH (Ki about 140 mM), shorter gamma-glutamylcysteine synthetase domain constructs are strongly inhibited (Ki about 15 mM) Streptococcus agalactiae
6.3.2.3 additional information not inhibited by glutathione Streptococcus agalactiae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.2.2 0.06
-
 ATP pH 8.4, 37°C, mutant enzyme D520stop Streptococcus agalactiae
6.3.2.2 0.065
-
 ATP pH 8.4, 37°C, mutant enzyme E494stop Streptococcus agalactiae
6.3.2.2 0.065
-
 ATP pH 8.4, 37°C, mutant enzyme G441stop Streptococcus agalactiae
6.3.2.2 0.066
-
 ATP pH 8.4, 37°C, mutant enzyme Y464stop Streptococcus agalactiae
6.3.2.2 0.069
-
 ATP pH 8.4, 37°C, mutant enzyme K526A Streptococcus agalactiae
6.3.2.2 0.082
-
 ATP pH 8.4, 37°C, mutant enzyme R508stop Streptococcus agalactiae
6.3.2.2 0.138
-
 L-Cys pH 8.4, 37°C, mutant enzyme Y464stop Streptococcus agalactiae
6.3.2.2 0.147
-
 L-Cys pH 8.4, 37°C, mutant enzyme K526A Streptococcus agalactiae
6.3.2.2 0.161
-
 L-Cys pH 8.4, 37°C, mutant enzyme D520stop Streptococcus agalactiae
6.3.2.2 0.166
-
 L-Cys pH 8.4, 37°C, mutant enzyme G441stop Streptococcus agalactiae
6.3.2.2 0.171
-
 L-Cys pH 8.4, 37°C, mutant enzyme E494stop Streptococcus agalactiae
6.3.2.2 0.26
-
 L-Cys pH 8.4, 37°C, mutant enzyme R508stop Streptococcus agalactiae
6.3.2.2 0.622
-
 ATP pH 8.4, 37°C, mutant enzyme H144A Streptococcus agalactiae
6.3.2.2 1
-
 L-Cys pH 8.4, 37°C, mutant enzyme H144A Streptococcus agalactiae
6.3.2.2 2 3 L-Glu pH 8.4, 37°C, mutant enzyme Y464stop Streptococcus agalactiae
6.3.2.2 22
-
 L-Glu pH 8.4, 37°C, mutant enzyme E494stop Streptococcus agalactiae
6.3.2.2 24
-
 L-Glu pH 8.4, 37°C, mutant enzyme K526A Streptococcus agalactiae
6.3.2.2 28.9
-
 L-Glu pH 8.4, 37°C, mutant enzyme D520stop Streptococcus agalactiae
6.3.2.2 34.7
-
 L-Glu pH 8.4, 37°C, mutant enzyme R508stop Streptococcus agalactiae
6.3.2.2 77
-
 L-Glu pH 8.4, 37°C, mutant enzyme G441stop Streptococcus agalactiae
6.3.2.2 229
-
 L-Glu pH 8.4, 37°C, mutant enzyme H144A Streptococcus agalactiae
6.3.2.3 0.42
-
 ATP wild type enzyme Streptococcus agalactiae
6.3.2.3 0.624
-
 ATP mutant enzyme K526A Streptococcus agalactiae
6.3.2.3 4.9
-
 ATP mutant enzyme D448A, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA Streptococcus agalactiae
6.3.2.3 5.9
-
 gamma-Glu-L-Cys wild type enzyme, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA Streptococcus agalactiae
6.3.2.3 6.3
-
 Gly wild type enzyme, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA Streptococcus agalactiae
6.3.2.3 7.5
-
 gamma-Glu-L-Cys mutant enzyme D448A, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA Streptococcus agalactiae
6.3.2.3 7.5
-
 gamma-Glu-L-Cys mutant enzyme K526A, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA Streptococcus agalactiae
6.3.2.3 11.8
-
 Gly mutant enzyme K526A, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA Streptococcus agalactiae
6.3.2.3 13.4
-
 ATP mutant enzyme K489A, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA Streptococcus agalactiae
6.3.2.3 23.8
-
 Gly mutant enzyme D448A, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA Streptococcus agalactiae
6.3.2.3 28
-
 Gly mutant enzyme K489A, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA Streptococcus agalactiae
6.3.2.3 31
-
 gamma-Glu-L-Cys mutant enzyme K489A, in 150 mM Tris-HCl buffer, pH 8.4, 100 mM KCl, 40 mM MgCl2, 0.3 mM EDTA Streptococcus agalactiae

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.3.2.3 88000
-
 2 * 88000, gel filtration Streptococcus agalactiae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.2.2 ATP + L-glutamate + L-cysteine Streptococcus agalactiae reaction is catalyzed by the bifunctional enzyme gamma-glutamylcysteine synthetase-glutathione synthetase ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.3.2.2 Streptococcus agalactiae
-
-
-
6.3.2.3 Streptococcus agalactiae
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.2.2
-
 Streptococcus agalactiae
6.3.2.3 Ni2+-NTA resin chromatography Streptococcus agalactiae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.2.2 ATP + L-glutamate + L-cysteine reaction is catalyzed by the bifunctional enzyme gamma-glutamylcysteine synthetase-glutathione synthetase Streptococcus agalactiae ADP + phosphate + gamma-L-glutamyl-L-cysteine
-
 ?
6.3.2.3 ATP + gamma-Glu-L-Cys + Gly
-
 Streptococcus agalactiae ADP + phosphate + glutathione
-
 ?

Subunits

EC Number Subunits Comment Organism
6.3.2.3 dimer 2 * 88000, gel filtration Streptococcus agalactiae

Synonyms

EC Number Synonyms Comment Organism
6.3.2.2 gamma-GCS
-
 Streptococcus agalactiae
6.3.2.2 gamma-glutamylcysteine synthetase-glutathione synthetase bifunctional enzyme Streptococcus agalactiae
6.3.2.3 gamma-GCS-GS bifunctional enzyme, with a glutathione synthetase at the C terminus Streptococcus agalactiae
6.3.2.3 gamma-glutamylcysteine synthetase-glutathione synthetase
-
 Streptococcus agalactiae

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.2.3 ATP
-
 Streptococcus agalactiae

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
6.3.2.2 7.6
-
 gamma-glutamylcysteine pH 8.4, 37°C, mutant enzyme Y464stop Streptococcus agalactiae
6.3.2.2 12.9
-
 GSH pH 8.4, 37°C, mutant enzyme Y464stop Streptococcus agalactiae
6.3.2.2 13.7
-
 gamma-glutamylcysteine pH 8.4, 37°C, mutant enzyme D520stop Streptococcus agalactiae
6.3.2.2 18.2
-
 GSH pH 8.4, 37°C, mutant enzyme E494stop Streptococcus agalactiae
6.3.2.2 23
-
 gamma-glutamylcysteine pH 8.4, 37°C, mutant enzyme R508stop Streptococcus agalactiae
6.3.2.2 24.3
-
 gamma-glutamylcysteine pH 8.4, 37°C, mutant enzyme E494stop Streptococcus agalactiae
6.3.2.2 277
-
 GSH pH 8.4, 37°C, mutant enzyme R508stop Streptococcus agalactiae
6.3.2.2 312
-
 GSH pH 8.4, 37°C, mutant enzyme D520stop Streptococcus agalactiae