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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

Cloned(Commentary)

Cloned (Comment) Organism
N-terminal His-tagged SySSADH expressed in Escherichia coli B834 (DE3) methionine auxotroph cells Synechococcus sp.

Crystallization (Commentary)

Crystallization (Comment) Organism
crystal structures of SySSADH determined in their apo form, as a binary complex with NADP+ and as a ternary complex with succinic semialdehyde and NADPH, resoultion of 1.7 A for the apo form and of 1.4 A for the binary and ternary complex Synechococcus sp.

Protein Variants

Protein Variants Comment Organism
C262A active site mutation, nonfunctional because Cys-262 acts as a nucleophilel Synechococcus sp.
E228A active site mutation, nonfunctional because Glu-228 acts as a general base Synechococcus sp.
E228Q active site mutation, nonfunctional because Glu-228 acts as a general base Synechococcus sp.
F132A activity of about 10–30% of the wild type enzyme, indicating a contribution of these succinic semialdehyde binding residues to the overall enzyme activity Synechococcus sp.
F425A inactive, suggesting that Phe-425 plays an important role in substrate binding Synechococcus sp.
I263A activity of about 10–30% of the wild type enzyme, indicating a contribution of these succinic semialdehyde binding residues to the overall enzyme activity Synechococcus sp.
N131A mutation of a residue that interacts with the O4 atom or the carboxyl group of succinic semialdehyde thus abolishing enzyme activity Synechococcus sp.
N131D mutation of a residue that interacts with the O4 atom or the carboxyl group of succinic semialdehyde thus abolishing enzyme activity Synechococcus sp.
R139A activity of about 10–30% of the wild type enzyme, indicating a contribution of these succinic semialdehyde binding residues to the overall enzyme activity Synechococcus sp.
R139K mutant enzyme exhibited an activity up to 80% that of the wild type enzyme, suggesting the significance of a positively charged residue in the binding of the carboxyl group of succinic semialdehyde Synechococcus sp.
S419A mutation of a residue that interacts with the O4 atom or the carboxyl group of succinic semialdehyde thus abolishing enzyme activity Synechococcus sp.
W135A activity of about 10–30% of the wild type enzyme, indicating a contribution of these succinic semialdehyde binding residues to the overall enzyme activity Synechococcus sp.

Inhibitors

Inhibitors Comment Organism Structure
H2O2 50 microM H2O2 sharply reduces activity to 31% of the H2O2-free enzyme and activity further decreases to 3% at 1 mM H2O2 Synechococcus sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information the Km-value of succinate semialdehyde estimated to be far less than 0.05 mM Synechococcus sp.
0.439
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NADP+ 30°C, pH 7.6 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. data from crystal structures provide details about the catalytic mechanism by revealing a covalent adduct of a cofactor with the catalytic cysteine in the binary complex and a proposed thiohemiacetal intermediate in the ternary complex succinate + NADPH + 2 H+
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Organism

Organism UniProt Comment Textmining
Synechococcus sp. B1XMM6
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Oxidation Stability

Oxidation Stability Organism
SySSADH is an oxidation-sensitive enzyme and the formation of the NADP-cysteine adduct is a kinetically preferred event that protects the catalytic Cys-262 from H2O2-dependent oxidative stress Synechococcus sp.

Purification (Commentary)

Purification (Comment) Organism
using immobilized metal affinity chromatography, removal of N-terminal His tag, further purification by immobilized metal affinity chromatography and gel filtration using a Superdex 200 column Synechococcus sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
succinate semialdehyde + NADP+ + H2O data from crystal structures provide details about the catalytic mechanism by revealing a covalent adduct of a cofactor with the catalytic cysteine in the binary complex and a proposed thiohemiacetal intermediate in the ternary complex Synechococcus sp. succinate + NADPH + 2 H+
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?

Subunits

Subunits Comment Organism
homodimer gel filtration, SySSADH monomer consists of three segments - alpha/beta-fold N- and C-domains for a cofactor binding and a catalytic domain, and three antiparallel beta-strands constituting a dimerization domain Synechococcus sp.

Synonyms

Synonyms Comment Organism
SSADH
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Synechococcus sp.
SySSADH
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Synechococcus sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5
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NADP+ 30°C, pH 7.6 Synechococcus sp.

General Information

General Information Comment Organism
metabolism succinic semialdehyde dehydrogenase from Synechococcus is an essential enzyme in the tricarboxylic acid cycle of cyanobacteria Synechococcus sp.