Cloned (Comment) | Organism |
---|---|
gene gsp, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
C59A | site-directed mutagenesis, inactive catalytic site mutant | Escherichia coli |
additional information | construction of a gene disruption mutant that does not produce glutathionylspermidine | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
45000 | - |
2 * 45000, SDS-PAGE, homodimer in which each monomer consists of two distinct domains. The C-terminal domain is responsible for the synthesis of glutathionylspermidine while the N-terminal domain catalyzes the hydrolysis of the conjugate | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
glutathionylspermidine + H2O | Escherichia coli | - |
glutathione + spermidine | GspSA catalyzes the formation of a peptide bond between GSH and spermidine, cf. EC 6.3.1.8 | r | |
additional information | Escherichia coli | bifunctional enzyme that catalyzes the ATP-dependent formation and hydrolysis of glutathionylspermidine, a conjugate of glutathione and spermidine. Proteins YgiC and YjfC, encoded by genes ygiC and yjfC, show ATPase activity, but are not glutathionylspermidine synthetases | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0AES0 | gene gsp | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by anion exchange chromatography and gel filtration | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
glutathionylspermidine + H2O | - |
Escherichia coli | glutathione + spermidine | GspSA catalyzes the formation of a peptide bond between GSH and spermidine, cf. EC 6.3.1.8 | r | |
glutathionylspermidine + H2O | the N-terminal domain catalyzes the hydrolysis of the conjugate | Escherichia coli | glutathione + spermidine | GspSA catalyzes the formation of a peptide bond between GSH and spermidine, cf. EC 6.3.1.8 | r | |
additional information | bifunctional enzyme that catalyzes the ATP-dependent formation and hydrolysis of glutathionylspermidine, a conjugate of glutathione and spermidine. Proteins YgiC and YjfC, encoded by genes ygiC and yjfC, show ATPase activity, but are not glutathionylspermidine synthetases | Escherichia coli | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 45000, SDS-PAGE, homodimer in which each monomer consists of two distinct domains. The C-terminal domain is responsible for the synthesis of glutathionylspermidine while the N-terminal domain catalyzes the hydrolysis of the conjugate | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
Glutathionylspermidine synthetase/amidase | - |
Escherichia coli |
GspSA | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
90 | - |
purified recombinant enzyme, pH 6.8, 3 min, inactivation | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.8 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | signature amino acid residues of ATP-grasp domain, R316, K498, K533, are responsible for the ATP binding | Escherichia coli |
General Information | Comment | Organism |
---|---|---|
evolution | the synthetase domain of GspSA belongs to the class of ATP-grasp structural domains. Proteins YgiC and YjfC proteins show 51% identity between themselves and 28% identity to the synthetase domain of the GspSA enzyme, but do not show any glutathionylspermidine synthetase/amidase enzyme activity | Escherichia coli |
additional information | comparison of catalytic properties of recombinant purified proteins GspSA, YgiC, and YjfC, overview | Escherichia coli |