Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 (DE3) cells | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapour diffusion method generating crystals of GspS in 0.1M TrisHCl (pH 8.5) containing 12% (v/v) PEG3350 and 0.5 M MgCl2 | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
C338A | increased Km compared to the wild type enzyme | Escherichia coli |
E391A | increased Km compared to the wild type enzyme | Escherichia coli |
E392A | increased Km compared to the wild type enzyme | Escherichia coli |
K607A | increased Km compared to the wild type enzyme | Escherichia coli |
R316E | no activity | Escherichia coli |
R538A | increased Km compared to the wild type enzyme | Escherichia coli |
R598A | decreased Km compared to the wild type enzyme | Escherichia coli |
S335A | increased Km compared to the wild type enzyme | Escherichia coli |
S337A | increased Km compared to the wild type enzyme | Escherichia coli |
T441A | increased Km compared to the wild type enzyme | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
(2-[[([2-[(4-amino-4-carboxybutanoyl)amino]propanoyl]amino)methyl](hydroxy)phosphoryl]ethyl)(hydroxy)dioxophosphate(1-) | inhibits the reaction in the presence of spermidine | Escherichia coli | |
4-[(14-ammonio-5-hydroxy-1-methyl-5-oxido-2-oxo-6-oxa-3,10-diaza-5-phosphatetradec-1-yl)amino]-1-carboxy-4-oxobutan-1-aminium | - |
Escherichia coli | |
4-[[2-([[[4-[(4-ammoniobutyl)amino]butyl](hydroxy)phosphoryl]methyl]amino)-1-methyl-2-oxoethyl]amino]-1-carboxy-4-oxobutan-1-aminium | - |
Escherichia coli | |
gamma-glutamyl-N-[[(3-aminopropoxy)(2-phosphonoethyl)phosphoryl]methyl]alaninamide | - |
Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0224 | - |
glutathione | mutant enzyme R598A | Escherichia coli | |
0.076 | - |
spermidine | wild type enzyme | Escherichia coli | |
0.076 | - |
spermidine | mutant enzyme S337A | Escherichia coli | |
0.126 | - |
spermidine | mutant enzyme S335A | Escherichia coli | |
0.153 | - |
spermidine | mutant enzyme K607A | Escherichia coli | |
0.182 | - |
spermidine | mutant enzyme R538A | Escherichia coli | |
0.218 | - |
glutathione | wild type enzyme | Escherichia coli | |
0.224 | - |
glutathione | mutant enzyme S337A | Escherichia coli | |
0.251 | - |
glutathione | mutant enzyme K607A | Escherichia coli | |
0.406 | - |
spermidine | mutant enzyme C338A | Escherichia coli | |
0.407 | - |
glutathione | mutant enzyme E391A | Escherichia coli | |
0.665 | - |
spermidine | mutant enzyme R598A | Escherichia coli | |
0.691 | - |
glutathione | mutant enzyme T441A | Escherichia coli | |
0.794 | - |
glutathione | mutant enzyme S335A | Escherichia coli | |
1.008 | - |
spermidine | mutant enzyme E392A | Escherichia coli | |
1.324 | - |
glutathione | mutant enzyme R538A | Escherichia coli | |
1.61 | - |
spermidine | mutant enzyme T441A | Escherichia coli | |
2.098 | - |
glutathione | mutant enzyme C338A | Escherichia coli | |
4.522 | - |
spermidine | mutant enzyme E391A | Escherichia coli | |
7.298 | - |
glutathione | mutant enzyme E392A | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | two Mg ions per molecule | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
70000 | - |
2 * 70000, analytical ultracentrifugation | Escherichia coli |
138000 | - |
analytical ultracentrifugation | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0AES0 | - |
- |
Purification (Comment) | Organism |
---|---|
nickel affinity column chromatography | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
glutathione + spermidine + ATP | - |
Escherichia coli | glutathionylspermidine + ADP + phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 70000, analytical ultracentrifugation | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
glutathionylspermidine synthetase | - |
Escherichia coli |
GspS | - |
Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Escherichia coli |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0000078 | - |
4-[[2-([[[4-[(4-ammoniobutyl)amino]butyl](hydroxy)phosphoryl]methyl]amino)-1-methyl-2-oxoethyl]amino]-1-carboxy-4-oxobutan-1-aminium | - |
Escherichia coli | |
0.006 | - |
4-[(14-ammonio-5-hydroxy-1-methyl-5-oxido-2-oxo-6-oxa-3,10-diaza-5-phosphatetradec-1-yl)amino]-1-carboxy-4-oxobutan-1-aminium | - |
Escherichia coli |