Cloned (Comment) | Organism |
---|---|
gene ssadh, recombinant expression in Escherichia coli strain BL21(DE3) | Streptococcus pyogenes |
Crystallization (Comment) | Organism |
---|---|
enzyme SpSSADH in a binary complex with succinate semialdehyde as the substrate and a ternary complex with succinate semialdehyde and NADP+, hanging-drop vapor diffusion method, mixing of mixture of 0.001 ml of protein solution with 0.001 ml of reservoir solution, for the binary complex crystal, SpSSADH is pre-incubated with succinate semialdehyde at the molar ratio of 1:2, and the protein-substrate mixture is crystallized over 00.5 ml of reservoir solution containing 0.1 M sodium acetate trihydrate, pH 4.6, and 2.0 M ammonium sulfate, the trinary complex is obtained by soaking the pre-grown NADP+ co-crystallized crystal with a 1:10 molar ratio of succinate semialdehyde under the same reservoir conditions, 22°C, X-ray diffraction structure determination and analysis at 2.4 A resolution, molecular replacement method with the apo-structure of SpSSADH, PDB ID 4OGD, as the search model | Streptococcus pyogenes |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | analysis of the kinetic inhibitory parameters revealed significant substrate inhibition in the presence of NADP+ at concentrations of succinate semialdehyde higher than 0.02 mM, which exhibits complete uncompetitive substrate inhibition, structure-based molecular insights into the substrate inhibition mechanism of SpSSADH, overview | Streptococcus pyogenes | |
succinate semialdehyde | complete uncompetitive substrate inhibition. Structural comparison of the tertiary enzyme-succinate semialdehyde-NADP+-complex with a binary complex of SpSSADH with NADP indicates that the substrate inhibition is induced by the binding of inhibitory succinate semialdehyde in the cofactor-binding site, instead of NADP+ | Streptococcus pyogenes |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Streptococcus pyogenes | |
0.00395 | - |
succinate semialdehyde | recombinant enzyme, pH 7.0, 30°C | Streptococcus pyogenes |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
succinate semialdehyde + NADP+ + H2O | Streptococcus pyogenes | - |
succinate + NADPH + 2 H+ | - |
? | |
succinate semialdehyde + NADP+ + H2O | Streptococcus pyogenes MG-AS1882 | - |
succinate + NADPH + 2 H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptococcus pyogenes | A0A0J9X1M8 | - |
- |
Streptococcus pyogenes MG-AS1882 | A0A0J9X1M8 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | in the binary enzyme-succinate semialdehyde-complex of SpSSADH, the succinate semialdehyde shows a tightly bound bent form nearby the catalytic residues, which may be caused by reduction of the cavity volume for substrate binding, compared with other SSADHs. Structural comparison of the tertiary enzyme-succinate semialdehyde-NADP+-complex with a binary complex + of SpSSADH with NADP indicates that the substrate inhibition is induced by the binding of inhibitory succinate semialdehyde in the cofactor-binding site, instead of NADP+. In the active site of enzyme SpSSADH, SSA is buried inside of the substrate-binding pocket formed by Phe133, Tyr136, Val262, Trp418 and Phe426 residues, substrate binding structure, overview | Streptococcus pyogenes | ? | - |
? | |
additional information | in the binary enzyme-succinate semialdehyde-complex of SpSSADH, the succinate semialdehyde shows a tightly bound bent form nearby the catalytic residues, which may be caused by reduction of the cavity volume for substrate binding, compared with other SSADHs. Structural comparison of the tertiary enzyme-succinate semialdehyde-NADP+-complex with a binary complex + of SpSSADH with NADP indicates that the substrate inhibition is induced by the binding of inhibitory succinate semialdehyde in the cofactor-binding site, instead of NADP+. In the active site of enzyme SpSSADH, SSA is buried inside of the substrate-binding pocket formed by Phe133, Tyr136, Val262, Trp418 and Phe426 residues, substrate binding structure, overview | Streptococcus pyogenes MG-AS1882 | ? | - |
? | |
succinate semialdehyde + NADP+ + H2O | - |
Streptococcus pyogenes | succinate + NADPH + 2 H+ | - |
? | |
succinate semialdehyde + NADP+ + H2O | - |
Streptococcus pyogenes MG-AS1882 | succinate + NADPH + 2 H+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
NADP+-dependent succinic semialdehyde dehydrogenase | - |
Streptococcus pyogenes |
SpSSADH | - |
Streptococcus pyogenes |
SSADH | - |
Streptococcus pyogenes |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Streptococcus pyogenes |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.69 | - |
succinate semialdehyde | recombinant enzyme, pH 7.0, 30°C | Streptococcus pyogenes |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Streptococcus pyogenes |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | cofactor binding structure, overview | Streptococcus pyogenes |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.1 | - |
succinate semialdehyde | recombinant enzyme, pH 7.0, 30°C | Streptococcus pyogenes |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
426 | - |
succinate semialdehyde | recombinant enzyme, pH 7.0, 30°C | Streptococcus pyogenes |