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Literature summary for 2.5.1.18 extracted from

  • Allocati, N.; Federici, L.; Masulli, M.; Favaloro, B.; Di Ilio, C.
    Cysteine 10 is critical for the activity of Ochrobactrum anthropi glutathione transferase and its mutation to alanine causes the preferential binding of glutathione to the H-site (2008), Proteins, 71, 16-23.
    View publication on PubMed
Search for more literature for 2.5.1.18

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli strain XL1-Blue Brucella anthropi

Crystallization (Commentary)

Crystallization (Comment) Organism
in the presence of glutathione, hanging drop vapour diffusion method, using 2 M ammonium sulfate as a precipitating agent Brucella anthropi

Protein Variants

Protein Variants Comment Organism
C10A the mutation causes the preferential binding of glutathione to the H-site, the mutant shows a decrease in activity of about 50%, drastic increase in Km value for glutathione of 105fold, and 23fold lower catalytic efficiency compared to the wild type enzyme Brucella anthropi
C10A/S11A mutant shows dramatic decrease in specific activity of about 98%, the double mutation exhibits loss of affinity for glutathione, a Km value 10-fold higher than in the wild type, and a 291fold decrease of the catalytic efficiency compared to the wild type enzyme Brucella anthropi

Inhibitors

Inhibitors Comment Organism Structure
guanidine hydrochloride retains 30% of its functionality at 0.5 M Brucella anthropi

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.132
-
 glutathione wild type enzyme, at 30°C Brucella anthropi
0.46
-
 1-chloro-2,4-dinitrobenzene mutant enzyme C10A/S11A, at 30°C Brucella anthropi
3.095
-
 1-chloro-2,4-dinitrobenzene wild type enzyme, at 30°C Brucella anthropi
3.933
-
 1-chloro-2,4-dinitrobenzene mutant enzyme C10A, at 30°C Brucella anthropi
13.93
-
 glutathione mutant enzyme C10A/S11A, at 30°C Brucella anthropi
13.96
-
 glutathione mutant enzyme C10A, at 30°C Brucella anthropi

Organism

Organism UniProt Comment Textmining
Brucella anthropi P81065
-
-

Purification (Commentary)

Purification (Comment) Organism
DEAE column chromatography and glutathione-affinity Sepharose column chromatography Brucella anthropi

Renatured (Commentary)

Renatured (Comment) Organism
in 0.1 M potassium phosphate buffer (pH 6.5) containing 1 mM EDTA and 5 mM dithiothreitol, with 4 M guanidine hydrochloride Brucella anthropi

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.08
-
 mutant enzyme C10A/S11A, at 30°C Brucella anthropi
3.1
-
 mutant enzyme C10A, at 30°C Brucella anthropi
5.9
-
 wild type enzyme, at 30°C Brucella anthropi

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
glutathione + 1-chloro-2,4-dinitrobenzene
-
 Brucella anthropi S-(2,4-dinitrophenyl)glutathione + HCl
-
 ?

Synonyms

Synonyms Comment Organism
glutathione S-transferase
-
 Brucella anthropi
GST
-
 Brucella anthropi

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
 wild type enzyme retains 80% of initial activity at about 50°C Brucella anthropi

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.45
-
 glutathione mutant enzyme C10A/S11A, at 30°C Brucella anthropi
0.85
-
 1-chloro-2,4-dinitrobenzene mutant enzyme C10A/S11A, at 30°C Brucella anthropi
2.35
-
 1-chloro-2,4-dinitrobenzene wild type enzyme, at 30°C Brucella anthropi
10.78
-
 1-chloro-2,4-dinitrobenzene mutant enzyme C10A, at 30°C Brucella anthropi
11.79
-
 glutathione mutant enzyme C10A, at 30°C Brucella anthropi
16.51
-
 glutathione wild type enzyme, at 30°C Brucella anthropi