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Literature summary for 6.3.2.2 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
Search for more literature for 6.3.2.2

Cloned(Commentary)

Cloned (Comment) Organism
-
 Streptococcus agalactiae

Protein Variants

Protein Variants Comment Organism
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
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
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
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
K526A KM-value for L-Glu, L-Cys and ATP are nearly identical to wild-type value Streptococcus agalactiae
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
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
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

Inhibitors

Inhibitors Comment Organism Structure
gamma-glutamylcysteine
-
 Streptococcus agalactiae
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

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.06
-
 ATP pH 8.4, 37°C, mutant enzyme D520stop Streptococcus agalactiae
0.065
-
 ATP pH 8.4, 37°C, mutant enzyme E494stop Streptococcus agalactiae
0.065
-
 ATP pH 8.4, 37°C, mutant enzyme G441stop Streptococcus agalactiae
0.066
-
 ATP pH 8.4, 37°C, mutant enzyme Y464stop Streptococcus agalactiae
0.069
-
 ATP pH 8.4, 37°C, mutant enzyme K526A Streptococcus agalactiae
0.082
-
 ATP pH 8.4, 37°C, mutant enzyme R508stop Streptococcus agalactiae
0.138
-
 L-Cys pH 8.4, 37°C, mutant enzyme Y464stop Streptococcus agalactiae
0.147
-
 L-Cys pH 8.4, 37°C, mutant enzyme K526A Streptococcus agalactiae
0.161
-
 L-Cys pH 8.4, 37°C, mutant enzyme D520stop Streptococcus agalactiae
0.166
-
 L-Cys pH 8.4, 37°C, mutant enzyme G441stop Streptococcus agalactiae
0.171
-
 L-Cys pH 8.4, 37°C, mutant enzyme E494stop Streptococcus agalactiae
0.26
-
 L-Cys pH 8.4, 37°C, mutant enzyme R508stop Streptococcus agalactiae
0.622
-
 ATP pH 8.4, 37°C, mutant enzyme H144A Streptococcus agalactiae
1
-
 L-Cys pH 8.4, 37°C, mutant enzyme H144A Streptococcus agalactiae
2 3 L-Glu pH 8.4, 37°C, mutant enzyme Y464stop Streptococcus agalactiae
22
-
 L-Glu pH 8.4, 37°C, mutant enzyme E494stop Streptococcus agalactiae
24
-
 L-Glu pH 8.4, 37°C, mutant enzyme K526A Streptococcus agalactiae
28.9
-
 L-Glu pH 8.4, 37°C, mutant enzyme D520stop Streptococcus agalactiae
34.7
-
 L-Glu pH 8.4, 37°C, mutant enzyme R508stop Streptococcus agalactiae
77
-
 L-Glu pH 8.4, 37°C, mutant enzyme G441stop Streptococcus agalactiae
229
-
 L-Glu pH 8.4, 37°C, mutant enzyme H144A Streptococcus agalactiae

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Streptococcus agalactiae
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Streptococcus agalactiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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
-
 ?

Synonyms

Synonyms Comment Organism
gamma-GCS
-
 Streptococcus agalactiae
gamma-glutamylcysteine synthetase-glutathione synthetase bifunctional enzyme Streptococcus agalactiae

Ki Value [mM]

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