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Literature summary for 6.3.2.3 extracted from

  • Merlino, A.; Russo Krauss, I.; Albino, A.; Pica, A.; Vergara, A.; Masullo, M.; De Vendittis, E.; Sica, F.
    Improving protein crystal quality by the without-oil microbatch method: crystallization and preliminary X-ray diffraction analysis of glutathione synthetase from Pseudoalteromonas haloplanktis (2011), Int. J. Mol. Sci., 12, 6312-6319.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
gene gshB, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) Pseudoalteromonas haloplanktis

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant enzyme, 20 mg/ml protein by hanging-drop method, from reservoir solution containing containing 10% w/v PEG 20K, 5% v/v tacsimate and 0.1 M HEPES, pH 7.5, 2-5 days, X-ray diffraction structure determination and analysis at 3.5 A resolution. Better crystals by vapor diffusion experiments to the without-oil microbatch method, method development, mixing of 40 mg/ml protein with an equal volume of solution containing 14% w/v PEG 20000, 0.2 M HEPES pH 7.5, 10% v/v tacsimate, X-ray diffraction structure determination and analysis at 2.34 A resolution, molecular replacement Pseudoalteromonas haloplanktis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Pseudoalteromonas haloplanktis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
36000
-
x * 36000, recombinant enzyme, SDS-PAGE Pseudoalteromonas haloplanktis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + gamma-L-glutamyl-L-cysteine + glycine Pseudoalteromonas haloplanktis
-
ADP + phosphate + glutathione
-
?

Organism

Organism UniProt Comment Textmining
Pseudoalteromonas haloplanktis
-
gene gshB
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Pseudoalteromonas haloplanktis

Reaction

Reaction Comment Organism Reaction ID
ATP + gamma-L-glutamyl-L-cysteine + glycine = ADP + phosphate + glutathione the reaction process involves the formation of an acyl phosphate on the cysteinyl moiety in L-gamma-glutamyl-L-cysteine, followed by the attack of the glycine and formation of an enzyme-product complex, which finally dissociates with the release of GSH, ADP and phosphate Pseudoalteromonas haloplanktis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + gamma-L-glutamyl-L-cysteine + glycine
-
Pseudoalteromonas haloplanktis ADP + phosphate + glutathione
-
?

Subunits

Subunits Comment Organism
? x * 36000, recombinant enzyme, SDS-PAGE Pseudoalteromonas haloplanktis

Synonyms

Synonyms Comment Organism
gshB
-
Pseudoalteromonas haloplanktis

Cofactor

Cofactor Comment Organism Structure
ATP
-
Pseudoalteromonas haloplanktis