Literature summary for 1.2.1.16 extracted from

Jang, E.H.; Park, S.A.; Chi, Y.M.; Lee, K.S.
Kinetic and structural characterization for cofactor preference of succinic semialdehyde dehydrogenase from Streptococcus pyogenes (2014), Mol. Cells, 37, 719-726 .

Cloned(Commentary)

Cloned (Comment)	Organism
gene ssadh, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)	Streptococcus pyogenes

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant SpSSADH in apo-form and in a binary complex with NADP+, hanging drop vapor diffusion method, for the apoform crystals: mixing 0.001 ml of protein solution with 0.001 ml of reservoir solution containing 0.1 M MES monohydrate, pH 5.8-6.1, and 18-23% w/v PEG 4000, and equilibration over 0.5 ml reservoir solution, for the binary complex crystals: mixing of protein solution with NADP+ in a 1:10 molar ratio with reservoir solution containing 0.1 M sodium acetate trihydrate, pH 4.6, and 2 M ammonium sulfate, 22°C, X-ray diffraction structure determination and analysis at 1.6 A and 2.1 A resolutions, respectively, molecular replacement method using the structure of Escherichia coli SSADH, PDB ID 3JZ4, as search model	Streptococcus pyogenes

Crystallization (Comment)

Organism

purified recombinant SpSSADH in apo-form and in a binary complex with NADP+, hanging drop vapor diffusion method, for the apoform crystals: mixing 0.001 ml of protein solution with 0.001 ml of reservoir solution containing 0.1 M MES monohydrate, pH 5.8-6.1, and 18-23% w/v PEG 4000, and equilibration over 0.5 ml reservoir solution, for the binary complex crystals: mixing of protein solution with NADP+ in a 1:10 molar ratio with reservoir solution containing 0.1 M sodium acetate trihydrate, pH 4.6, and 2 M ammonium sulfate, 22°C, X-ray diffraction structure determination and analysis at 1.6 A and 2.1 A resolutions, respectively, molecular replacement method using the structure of Escherichia coli SSADH, PDB ID 3JZ4, as search model

Streptococcus pyogenes

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
additional information	-	additional information	Michaelis-Menten kinetics, kinetic analysis	Streptococcus pyogenes
0.1	-	NADP+	recombinant His6-tagged enzyme, pH 7.0, 30°C	Streptococcus pyogenes
0.23	-	NAD+	recombinant His6-tagged 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+	-	?

Organism

Organism	UniProt	Comment	Textmining
Streptococcus pyogenes	A0A0J9X1M8	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration	Streptococcus pyogenes

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
succinate semialdehyde + NAD+ + H2O	-	Streptococcus pyogenes	succinate + NADH + 2 H+	-	?
succinate semialdehyde + NADP+ + H2O	-	Streptococcus pyogenes	succinate + NADPH + 2 H+	-	?

Subunits

Subunits	Comment	Organism
homodimer	enzyme SpSSADH shows a dimeric conformation, containing a single cysteine residue in the catalytic loop of each subunit	Streptococcus pyogenes

Synonyms

Synonyms	Comment	Organism
SpSSADH	-	Streptococcus pyogenes
SSADH	-	Streptococcus pyogenes
succinic semialdehyde dehydrogenase	-	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
1.94	-	NAD+	recombinant His6-tagged enzyme, pH 7.0, 30°C	Streptococcus pyogenes
7.2	-	NADP+	recombinant His6-tagged 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
NAD+	-	Streptococcus pyogenes
NADP+	SpSSADH prefers NADP+ over NAD+ as a hydride acceptor, structural basis of the cofactor preference of SpSSADH, overview. Residues Ser158 and Tyr188 in SpSSADH participate in the stabilization of the 2'-phosphate group of adenine-side ribose in NADP+	Streptococcus pyogenes

General Information

General Information	Comment	Organism
metabolism	succinic semialdehyde dehydrogenase (SSADH) catalyzes the oxidation of succinic semialdehyde to succinic acid in the final step of GABA catabolism	Streptococcus pyogenes

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
9.7	-	NAD+	recombinant His6-tagged enzyme, pH 7.0, 30°C	Streptococcus pyogenes
72	-	NADP+	recombinant His6-tagged enzyme, pH 7.0, 30°C	Streptococcus pyogenes