Cloned (Comment) | Organism |
---|---|
gene gshB, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Pseudoalteromonas haloplanktis |
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 | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Pseudoalteromonas haloplanktis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
36000 | - |
x * 36000, recombinant enzyme, SDS-PAGE | Pseudoalteromonas haloplanktis |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Pseudoalteromonas haloplanktis | - |
gene gshB | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Pseudoalteromonas haloplanktis |
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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + gamma-L-glutamyl-L-cysteine + glycine | - |
Pseudoalteromonas haloplanktis | ADP + phosphate + glutathione | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 36000, recombinant enzyme, SDS-PAGE | Pseudoalteromonas haloplanktis |
Synonyms | Comment | Organism |
---|---|---|
gshB | - |
Pseudoalteromonas haloplanktis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Pseudoalteromonas haloplanktis |