Literature summary 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)
EC Number |
Cloned (Comment) |
Organism |
---|
1.2.1.79 |
recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21/DE3 |
Synechococcus sp. PCC 7002 |
Crystallization (Commentary)
EC Number |
Crystallization (Comment) |
Organism |
---|
1.2.1.79 |
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
EC Number |
Protein Variants |
Comment |
Organism |
---|
1.2.1.79 |
C262A |
site-directed mutagenesis, Sp2771 mutant structure analysis and comparison to the wild-type structure |
Synechococcus sp. PCC 7002 |
1.2.1.79 |
S419A |
site-directed mutagenesis, Sp2771 mutant structure analysis and comparison to the wild-type structure |
Synechococcus sp. PCC 7002 |
Natural Substrates/ Products (Substrates)
EC Number |
Natural Substrates |
Organism |
Comment (Nat. Sub.) |
Natural Products |
Comment (Nat. Pro.) |
Rev. |
Reac. |
---|
1.2.1.79 |
succinate semialdehyde + NADP+ + H2O |
Synechococcus sp. PCC 7002 |
- |
succinate + NADPH + 2 H+ |
- |
? |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
1.2.1.79 |
Synechococcus sp. PCC 7002 |
B1XMM6 |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
1.2.1.79 |
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)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
1.2.1.79 |
succinate semialdehyde + NADP+ + H2O |
- |
Synechococcus sp. PCC 7002 |
succinate + NADPH + 2 H+ |
- |
? |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
1.2.1.79 |
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
EC Number |
Synonyms |
Comment |
Organism |
---|
1.2.1.79 |
Sp2771 |
- |
Synechococcus sp. PCC 7002 |
1.2.1.79 |
SSADH |
- |
Synechococcus sp. PCC 7002 |
1.2.1.79 |
succinic semialdehy de dehydrogenase |
- |
Synechococcus sp. PCC 7002 |
1.2.1.79 |
SYNPCC7002_A2771 |
gene name, UniProt |
Synechococcus sp. PCC 7002 |
Temperature Optimum [°C]
EC Number |
Temperature Optimum [°C] |
Temperature Optimum Maximum [°C] |
Comment |
Organism |
---|
1.2.1.79 |
22 |
- |
assay at room temperature |
Synechococcus sp. PCC 7002 |
Cofactor
EC Number |
Cofactor |
Comment |
Organism |
Structure |
---|
1.2.1.79 |
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
EC Number |
General Information |
Comment |
Organism |
---|
1.2.1.79 |
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 |