Literature summary for 1.2.1.79 extracted from

Yuan, Z.; Yin, B.; Wei, D.; Yuan, Y.R.
Structural basis for cofactor and substrate selection by cyanobacterium succinic semialdehyde dehydrogenase (2013), J. Struct. Biol., 182, 125-135 .

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21/DE3	Synechococcus sp. PCC 7002

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His6-tagged wild-type Sp2771, and Sp2771 S419A and Sp2771 C262A mutants in ternary complex with NADP+ and succinate semialdehyde, mixing of 0.001 ml of protein solution and reservoir solution each, the latter containing 15% PEG 5000 MME , 1 mM DTT, 3% tascimate, and 100 mM HEPES, pH 6.8 for the wild-type enzyme and the mutants, for Sp2771 mutants in complex with NADP+ and succinate semialdehyde, NAD+ and succinate semialdehyde are incubated with proteins for 15 min at room temperature before crystallization, 20°C, 3 days, X-ray diffraction structure determination and analysis at 1.7-2.5 A resolution, molecular replacement using Escherichia coli SSADH structure, PDB ID 3JZ4, as search model	Synechococcus sp. PCC 7002

Crystallization (Comment)

Organism

purified recombinant His6-tagged wild-type Sp2771, and Sp2771 S419A and Sp2771 C262A mutants in ternary complex with NADP+ and succinate semialdehyde, mixing of 0.001 ml of protein solution and reservoir solution each, the latter containing 15% PEG 5000 MME , 1 mM DTT, 3% tascimate, and 100 mM HEPES, pH 6.8 for the wild-type enzyme and the mutants, for Sp2771 mutants in complex with NADP+ and succinate semialdehyde, NAD+ and succinate semialdehyde are incubated with proteins for 15 min at room temperature before crystallization, 20°C, 3 days, X-ray diffraction structure determination and analysis at 1.7-2.5 A resolution, molecular replacement using Escherichia coli SSADH structure, PDB ID 3JZ4, as search model

Synechococcus sp. PCC 7002

Protein Variants

Protein Variants	Comment	Organism
C262A	site-directed mutagenesis, Sp2771 mutant structure analysis and comparison to the wild-type structure	Synechococcus sp. PCC 7002
S419A	site-directed mutagenesis, Sp2771 mutant structure analysis and comparison to the wild-type structure	Synechococcus sp. PCC 7002

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
succinate semialdehyde + NADP+ + H2O	Synechococcus sp. PCC 7002	-	succinate + NADPH + 2 H+	-	?

Organism

Organism	UniProt	Comment	Textmining
Synechococcus sp. PCC 7002	B1XMM6	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, gel filtration, and ultrafiltration	Synechococcus sp. PCC 7002

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
succinate semialdehyde + NADP+ + H2O	-	Synechococcus sp. PCC 7002	succinate + NADPH + 2 H+	-	?

Subunits

Subunits	Comment	Organism
More	the central parts of both cofactor binding domain and catalytic domain contain atypical alpha/beta structure. The catalytic domain consists of a seven stranded beta-sheet flanked by two alpha-helices on one side and three alpha-helices on the other side.The cofactor binding domain displays the two tandem Rossmann folds for NADP+ binding . The oligomerization domain contains three-stranded antiparallel beta-sheets protruding across the center of the dimer interface, while the NADP+ binding site is on the opposite side of the dimer interface	Synechococcus sp. PCC 7002

Synonyms

Synonyms	Comment	Organism
Sp2771	-	Synechococcus sp. PCC 7002
SSADH	-	Synechococcus sp. PCC 7002
succinic semialdehy de dehydrogenase	-	Synechococcus sp. PCC 7002
SYNPCC7002_A2771	gene name, UniProt	Synechococcus sp. PCC 7002

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
22	-	assay at room temperature	Synechococcus sp. PCC 7002

Cofactor

Cofactor	Comment	Organism	Structure
NADP+	Ser157 residue in Sp2771 plays a critical structural role in determining NADP+ preference for Sp2771, whereas size and distribution of hydrophobic residues along the substrate binding funnel determine substrate selection	Synechococcus sp. PCC 7002

General Information

General Information	Comment	Organism
additional information	Ser157 residue in Sp2771 plays a critical structural role in determining NADP+ preference for Sp2771, whereas size and distribution of hydrophobic residues along the substrate binding funnel determine substrate selection. Enzyme Sp2771 structure modelling comprising residues 2-454, active site and substrate binding structures, overview	Synechococcus sp. PCC 7002