Literature summary for 1.2.1.79 extracted from

Jang, E.H.; Park, S.A.; Chi, Y.M.; Lee, K.S.
Structural insight into the substrate inhibition mechanism of NADP+-dependent succinic semialdehyde dehydrogenase from Streptococcus pyogenes (2015), Biochem. Biophys. Res. Commun., 461, 487-493 .

Cloned(Commentary)

Cloned (Comment)	Organism
gene ssadh, recombinant expression in Escherichia coli strain BL21(DE3)	Streptococcus pyogenes

Crystallization (Commentary)

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

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

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 [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/ Products (Substrates)

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

Organism	UniProt	Comment	Textmining
Streptococcus pyogenes	A0A0J9X1M8	-	-
Streptococcus pyogenes MG-AS1882	A0A0J9X1M8	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
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

Synonyms	Comment	Organism
NADP+-dependent succinic semialdehyde dehydrogenase	-	Streptococcus pyogenes
SpSSADH	-	Streptococcus pyogenes
SSADH	-	Streptococcus pyogenes

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Streptococcus pyogenes

Turnover Number [1/s]

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

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	assay at	Streptococcus pyogenes

Cofactor

Cofactor	Comment	Organism	Structure
NADP+	cofactor binding structure, overview	Streptococcus pyogenes

Ki Value [mM]

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 [mM/s]

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