Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.2.1.79 extracted from

  • Park, J.; Rhee, S.
    Structural basis for a cofactor-dependent oxidation protection and catalysis of cyanobacterial succinic semialdehyde dehydrogenase (2013), J. Biol. Chem., 288, 15760-15770 .
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
DTT enzyme activity increases significantly, and the enzyme becomes resistant to oxidative stress in presence of NADP+ and DTT Synechococcus sp.

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of N-terminally His-tagged SySSADH (residues Met1 to Lys454) in Escherichia coli methionine-auxotrophic strain B834(DE3), recombinant expression of His-tagged enzyme mutants in Escherichia coli strain coli BL21(DE3). Juxtaposition of the N- and C-domains generates an active site tunnel between the two domains that is accessible from both ends. The catalytic residues are located in the middle of the tunnel. A nucleophile Cys262 is located in the catalytic loop between alphaa8 and beta11 of the C-domain, and a general base Glu228 is located in an interdomain-connecting loop between beta9 and beta10. Dimerization is mediated largely by N-domain alpha7 and the three antiparallel beta-strands (beta3, beta4, beta17) protruding from the N- and C-domains Synechococcus sp.

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant enzyme in apo form, in a binary complex with NADP+, and in a ternary complex with succinic semialdehyde and NADPH, sitting drop vapor diffusion method, using a crystallization buffer of 0.05 M potassium phosphate monobasic, 20% w/v PEG 8000, and 2 mM CaCl2, 22°C, for the binary and tertiary complexes, a pre-grown crystals of SySSADH are soaked for 60 min in a solution of 0.05 M potassium phosphate monobasic, 20% w/v PEG8000, 30% v/v ethylene glycol, and 50 mM NADPH or 50 mM NADPH and 50 mM succinate semialdehyde, respectively, X-ray diffraction structure determination and analysis at 1.4-1.7 A resolution, single-wavelength diffraction, modelling Synechococcus sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
succinate semialdehyde + NADP+ + H2O Synechococcus sp.
-
succinate + NADPH + 2 H+
-
r
succinate semialdehyde + NADP+ + H2O Synechococcus sp. ATCC 27264
-
succinate + NADPH + 2 H+
-
r

Organism

Organism UniProt Comment Textmining
Synechococcus sp. B1XMM6
-
-
Synechococcus sp. ATCC 27264 B1XMM6
-
-

Oxidation Stability

Oxidation Stability Organism
SySSADH is an oxidation-sensitive enzyme, the formation of the NADP-cysteine adduct is a kinetically preferred event that protects the catalytic cysteine from H2O2-dependent oxidative stress. Over 70% of the original SySSADH activity is maintained with 0.005-0.25 mM H2O2 with a further drop of activity to 40% at 1 m H2O2. Comparable or even higher enzyme activity is observed when SySSADH is preincubated for 10 min with 2.5 mM NADP+ followed by H2O2 treatment for 60 min Synechococcus sp.

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminally His-tagged SySSADH from Escherichia coli strain B834(DE3) by nickel affinity chromatography, dialysis and cleavge fo the His-tag by tobacco etch mosaic virus protease, followed by another step of immobilized metal affinity chromatography and gel filtration. Recombinant His-tagged enzyme mutants from Escherichia coli strain coli BL21(DE3) by immobilized metal affinity chromatography and dialysis Synechococcus sp.

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
39.6
-
purified recombinant enzyme, pH 7.6, 30°C Synechococcus sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
succinate semialdehyde + NADP+ + H2O
-
Synechococcus sp. succinate + NADPH + 2 H+
-
r
succinate semialdehyde + NADP+ + H2O binding structure, overview Synechococcus sp. succinate + NADPH + 2 H+
-
r
succinate semialdehyde + NADP+ + H2O
-
Synechococcus sp. ATCC 27264 succinate + NADPH + 2 H+
-
r
succinate semialdehyde + NADP+ + H2O binding structure, overview Synechococcus sp. ATCC 27264 succinate + NADPH + 2 H+
-
r

Subunits

Subunits Comment Organism
homodimer the overall structure of monomeric SySSADH is reminiscent of an ALDH fold. It is made up of three segments: alpha/beta-fold N- and C-domains for a cofactor binding and a catalytic domain, respectively, and three antiparallel beta-strands constituting a dimerization domain Synechococcus sp.

Synonyms

Synonyms Comment Organism
SSADH
-
Synechococcus sp.
succinic semialdehyde dehydrogenase
-
Synechococcus sp.
SySSADH
-
Synechococcus sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Synechococcus sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
assay at Synechococcus sp.

Cofactor

Cofactor Comment Organism Structure
NADP+ enzyme activity increases significantly, and the enzyme becomes resistant to oxidative stress in presence of NADP+ and DTT Synechococcus sp.
NADPH binding structure, overview Synechococcus sp.

General Information

General Information Comment Organism
evolution SSADH belongs to the aldehyde dehydrogenase (ALDH) superfamily Synechococcus sp.
additional information structure analysis of the enzyme in binary and ternary with NADP(H) and/or substrate reveals that the enzyme forms a distinct reaction intermediate in each complex: a covalent adduct of a cofactor with the catalytic cysteine in the binary complex and a proposed thiohemiacetal intermediate in the ternary complex. SySSADH produces succinate in an NADP+ -dependent manner with a single cysteine acting as the catalytic residue in the catalytic loop, catalytic mechanism, overview. The formation of the NADP-cysteine adduct is a kinetically preferred event that protects the catalytic cysteine from H2O2-dependent oxidative stress. SySSADH shows a cofactor-dependent oxidation protection in 1-Cys SSADH, which is unique relative to other 2-Cys SSADHs employing a redox-dependent formation of a disulfide bridge. The catalytic cysteine preferentially forms an NADP-cysteine adduct if NADP+ is present Synechococcus sp.
physiological function succinic semialdehyde dehydrogenase from Synechococcus is an essential enzyme in the tricarboxylic acid, TCA, cycle of cyanobacteria. It completes a 2-oxoglutarate dehydrogenase-deficient cyanobacterial TCA cycle through a detour metabolic pathway. SySSADH produces succinate in an NADP+ -dependent manner with a single cysteine acting as the catalytic residue in the catalytic loop Synechococcus sp.