Cloned (Comment) | Organism |
---|---|
expression of enzyme in Escherichia coli strain BL21(DE3) | Synechocystis sp. |
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3). K273A mutant forms inclusion bodies | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme, hanging drop method, mixing of protein solution containing 10 mg/ml protein in 10 mM NaHCO3, 10 mM 1-hydroxy-2-naphthoyl-CoA, and 25 mM Tris, pH 8.0, with reservoir solution containing 200 mM (NH4)2SO4 and 23% PEG 3,350 in 100 mM Bis-Tris, pH 5.5, in a 1:1 ratio, 1 week, X-ray diffraction structure determination and analysis at 1.84 A resolution | Escherichia coli |
purified recombinant enzyme, hanging drop method, mixing of protein solution containing 10 mg/ml protein in 10 mM NaHCO3, 5 mM 1-hydroxy-2-naphthoyl-CoA, 5 mM salicyloyl-CoA, 20 mM glycine, pH 9.75, 1% glycerol, and 10 mM NaHCO3 or 0.15 M ammonium acetate, 4% tacsimate, 15% PEG 3350, and 100 mM Bis-Tris, pH 6.0, with reservoir solution containing 0.15 M ammonium acetate, 0.3 M ammonium sulfate, 16% PEG 3350, 100 mM Bis-Tris, pH 5.7, and 10 mM proline or 10 mM taurine, in a 1:1 ratio, 1 week, X-ray diffraction structure determination and analysis at 2.0-2.35 A resolution | Synechocystis sp. |
Protein Variants | Comment | Organism |
---|---|---|
F270A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Escherichia coli |
K89A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Escherichia coli |
K91A | site-directed mutagenesis, inactive mutant | Escherichia coli |
R267A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
1-hydroxy-2-naphthoyl-CoA | a product analogue, protein-ligand interactions, complex structure, overview | Escherichia coli | |
1-hydroxy-2-naphthoyl-CoA | a product analogue, protein-ligand interactions, complex structure, overview | Synechocystis sp. | |
salicyloyl-CoA | protein-ligand interactions, complex structure, overview | Escherichia coli | |
salicyloyl-CoA | protein-ligand interactions, complex structure, overview | Synechocystis sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0028 | - |
o-succinylbenzoate | pH 7.0, temperature not specified in the publication, wild-type enzyme | Escherichia coli | |
0.0166 | - |
o-succinylbenzoate | pH 7.0, temperature not specified in the publication, mutant R267A | Escherichia coli | |
0.0197 | - |
o-succinylbenzoate | pH 7.0, temperature not specified in the publication, mutant F270A | Escherichia coli | |
0.0433 | - |
o-succinylbenzoate | pH 7.0, temperature not specified in the publication, mutant K89A | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
o-succinylbenzoate + CoA | Synechocystis sp. | formation of a structural motif like an oxyanion hole for orientation of the o-succinylbenzoate substrate and stabilization of the oxyanion intermediate in the intramolecular Claisen condensation, which involves a conserved tyrosine residue in the A-loop via induced fit | 1,4-dihydroxy-2-naphthoyl-CoA + H2O | - |
? | |
o-succinylbenzoate + CoA | Escherichia coli | formation of a structural motif like an oxyanion hole for orientation of the o-succinylbenzoate substrate and stabilization of the oxyanion intermediate in the intramolecular Claisen condensation, which involves a conserved tyrosine residue in the A-loop. via induced fit | 1,4-dihydroxy-2-naphthoyl-CoA + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Synechocystis sp. | P73495 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme from Escherichia coli strain BL21(DE3) too over 95% purity | Synechocystis sp. |
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21 (DE3) too over 95% purity | Escherichia coli |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
4-(2-carboxyphenyl)-4-oxobutanoyl-CoA = 1,4-dihydroxy-2-naphthoyl-CoA + H2O | unique induced-fit catalytic mechanism which involves intersubunit interactions, overview | Escherichia coli | |
4-(2-carboxyphenyl)-4-oxobutanoyl-CoA = 1,4-dihydroxy-2-naphthoyl-CoA + H2O | unique induced-fit catalytic mechanism which involves intersubunit interactions, overview | Synechocystis sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
o-succinylbenzoate + CoA | - |
Escherichia coli | 1,4-dihydroxy-2-naphthoyl-CoA + H2O | - |
? | |
o-succinylbenzoate + CoA | - |
Synechocystis sp. | 1,4-dihydroxy-2-naphthoyl-CoA + H2O | - |
? | |
o-succinylbenzoate + CoA | formation of a structural motif like an oxyanion hole for orientation of the o-succinylbenzoate substrate and stabilization of the oxyanion intermediate in the intramolecular Claisen condensation, which involves a conserved tyrosine residue in the A-loop via induced fit | Synechocystis sp. | 1,4-dihydroxy-2-naphthoyl-CoA + H2O | - |
? | |
o-succinylbenzoate + CoA | formation of a structural motif like an oxyanion hole for orientation of the o-succinylbenzoate substrate and stabilization of the oxyanion intermediate in the intramolecular Claisen condensation, which involves a conserved tyrosine residue in the A-loop. via induced fit | Escherichia coli | 1,4-dihydroxy-2-naphthoyl-CoA + H2O | - |
? |
Synonyms | Comment | Organism |
---|---|---|
1,4-dihydroxy-2-naphthoyl coenzyme A synthase | - |
Escherichia coli |
1,4-dihydroxy-2-naphthoyl coenzyme A synthase | - |
Synechocystis sp. |
DHNA-CoA synthase | - |
Escherichia coli |
DHNA-CoA synthase | - |
Synechocystis sp. |
MenB | - |
Escherichia coli |
MenB | - |
Synechocystis sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0027 | - |
o-succinylbenzoate | pH 7.0, temperature not specified in the publication, mutant R267A | Escherichia coli | |
0.0035 | - |
o-succinylbenzoate | pH 7.0, temperature not specified in the publication, mutant F270A | Escherichia coli | |
0.021 | - |
o-succinylbenzoate | pH 7.0, temperature not specified in the publication, wild-type enzyme | Escherichia coli | |
0.038 | - |
o-succinylbenzoate | pH 7.0, temperature not specified in the publication, mutant K89A | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Escherichia coli |
7 | - |
assay at | Synechocystis sp. |
General Information | Comment | Organism |
---|---|---|
metabolism | the enzyme is involved in the menaquinone biosynthesis | Escherichia coli |
metabolism | the enzyme is involved in the menaquinone biosynthesis | Synechocystis sp. |
additional information | enzyme structure in complex with a product analogue, the structural changes include the folding of an active-site loop into a beta-hairpin and significant reorientation of a helix at the carboxy terminus. A new interface is formed between the ordered loop and the reoriented helix, both of which also form additional interactions with the coenzyme A moiety of the ligand | Synechocystis sp. |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.16 | - |
o-succinylbenzoate | pH 7.0, temperature not specified in the publication, mutant R267A | Escherichia coli | |
0.18 | - |
o-succinylbenzoate | pH 7.0, temperature not specified in the publication, mutant F270A | Escherichia coli | |
0.89 | - |
o-succinylbenzoate | pH 7.0, temperature not specified in the publication, mutant K89A | Escherichia coli | |
7.4 | - |
o-succinylbenzoate | pH 7.0, temperature not specified in the publication, wild-type enzyme | Escherichia coli |