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Literature summary for 1.1.1.25 extracted from

  • Singh, S.A.; Christendat, D.
    Structure of Arabidopsis dehydroquinate dehydratase-shikimate dehydrogenase and implications for metabolic channeling in the shikimate pathway (2006), Biochemistry, 45, 10406.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of C-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Arabidopsis thaliana

Crystallization (Commentary)

Crystallization (Comment) Organism
purified native and selenomethionine-labeled DELTA88DHQ-SDH complexed with shikimate, hanging drop vapour diffusion method, 10 mg/ml protein in 1 mM shikimate mixed with equal volume of reservoir solution containing 0.4 M potassium sodium tartrate tetrahydrate, 0.1 M tri-sodium dihydrate, pH 5.6, and 2.8 M ammonium sulfate, X-ray diffraction structure determination and analysi at 1.95-2.2 A resolution Arabidopsis thaliana

Protein Variants

Protein Variants Comment Organism
D423A site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, inactive mutant Arabidopsis thaliana
D423N site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, highly reduced activity compared to the wild-type enzyme Arabidopsis thaliana
K385A site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, highly reduced activity compared to the wild-type enzyme Arabidopsis thaliana
K385N site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, highly reduced activity compared to the wild-type enzyme Arabidopsis thaliana
K385N/D423N site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, highly reduced activity compared to the wild-type enzyme Arabidopsis thaliana
additional information construction of deletion variant DELTA88DHQ-SDH, thermal denaturation analysis, overview Arabidopsis thaliana
S336A site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, reduced activity compared to the wild-type enzyme Arabidopsis thaliana
S338A site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, reduced activity compared to the wild-type enzyme Arabidopsis thaliana
Y550A site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, reduced activity compared to the wild-type enzyme Arabidopsis thaliana
Y550F site-directed mutagenesis of enzyme variant DELTA88DHQ-SDH, reduced activity compared to the wild-type enzyme Arabidopsis thaliana

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.017
-
NADP+ pH 7.5, 22°C, mutant K385A Arabidopsis thaliana
0.022
-
NADP+ pH 7.5, 22°C, mutant K385N/D423N Arabidopsis thaliana
0.053
-
NADP+ pH 7.5, 22°C, mutant K385N Arabidopsis thaliana
0.079
-
NADP+ pH 7.5, 22°C, mutant D423N Arabidopsis thaliana
0.114
-
NADP+ pH 7.5, 22°C, mutant S338A Arabidopsis thaliana
0.131
-
shikimate pH 7.5, 22°C, mutant K385N/D423N Arabidopsis thaliana
0.131
-
NADP+ pH 7.5, 22°C, wild-type DELTA88DHQ-SDH enzyme variant Arabidopsis thaliana
0.132
-
NADP+ pH 7.5, 22°C, mutant Y550F Arabidopsis thaliana
0.152
-
NADP+ pH 7.5, 22°C, mutant S336A Arabidopsis thaliana
0.157
-
NADP+ pH 7.5, 22°C, mutant Y550A Arabidopsis thaliana
0.263
-
shikimate pH 7.5, 22°C, mutant K385A Arabidopsis thaliana
0.47
-
shikimate pH 7.5, 22°C, mutant K385N Arabidopsis thaliana
0.555
-
shikimate pH 7.5, 22°C, mutant D423N Arabidopsis thaliana
0.685
-
shikimate pH 7.5, 22°C, wild-type DELTA88DHQ-SDH enzyme variant Arabidopsis thaliana
1.03
-
shikimate pH 7.5, 22°C, mutant Y550A Arabidopsis thaliana
1.06
-
shikimate pH 7.5, 22°C, mutant Y550F Arabidopsis thaliana
6.2
-
shikimate pH 7.5, 22°C, mutant S338A Arabidopsis thaliana
8.74
-
shikimate pH 7.5, 22°C, mutant S336A Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3-dehydroshikimate + NADPH Arabidopsis thaliana
-
shikimate + NADP+
-
r
additional information Arabidopsis thaliana the bifunctional enzyme performs dehydroquinate dehydratase and shikimate dehydrogenase activities, metabolic channeling ?
-
?

Organism

Organism UniProt Comment Textmining
Arabidopsis thaliana
-
bifunctional enzyme
-

Reaction

Reaction Comment Organism Reaction ID
shikimate + NADP+ = 3-dehydroshikimate + NADPH + H+ active site structure, reaction and kinetic mechanisms Arabidopsis thaliana

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Arabidopsis thaliana

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3-dehydroshikimate + NADPH
-
Arabidopsis thaliana shikimate + NADP+
-
r
additional information the bifunctional enzyme performs dehydroquinate dehydratase and shikimate dehydrogenase activities, metabolic channeling Arabidopsis thaliana ?
-
?
additional information the bifunctional enzyme performs dehydroquinate dehydratase and shikimate dehydrogenase activities Arabidopsis thaliana ?
-
?

Synonyms

Synonyms Comment Organism
dehydroquinate dehydratase-shikimate dehydrogenase
-
Arabidopsis thaliana
DHQ-SDH
-
Arabidopsis thaliana
SDH
-
Arabidopsis thaliana
shikimate dehydrogenase
-
Arabidopsis thaliana

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
-
assay at Arabidopsis thaliana

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.008
-
NADP+ pH 7.5, 22°C, mutant K385A Arabidopsis thaliana
0.013
-
shikimate pH 7.5, 22°C, mutant K385A Arabidopsis thaliana
0.036
-
NADP+ pH 7.5, 22°C, mutant K385N/D423N Arabidopsis thaliana
0.043
-
shikimate pH 7.5, 22°C, mutant K385N/D423N Arabidopsis thaliana
0.063
-
shikimate pH 7.5, 22°C, mutant K385N Arabidopsis thaliana
0.066
-
NADP+ pH 7.5, 22°C, mutant K385N Arabidopsis thaliana
0.092
-
NADP+ pH 7.5, 22°C, mutant D423N Arabidopsis thaliana
0.121
-
shikimate pH 7.5, 22°C, mutant D423N Arabidopsis thaliana
1.86
-
NADP+ pH 7.5, 22°C, mutant Y550A Arabidopsis thaliana
2.1
-
shikimate pH 7.5, 22°C, mutant Y550A Arabidopsis thaliana
3.11
-
NADP+ pH 7.5, 22°C, mutant Y550F Arabidopsis thaliana
3.9
-
shikimate pH 7.5, 22°C, mutant Y550F Arabidopsis thaliana
11.2
-
NADP+ pH 7.5, 22°C, mutant S338A Arabidopsis thaliana
33.2
-
shikimate pH 7.5, 22°C, mutant S338A Arabidopsis thaliana
52.9
-
NADP+ pH 7.5, 22°C, mutant S336A Arabidopsis thaliana
140
-
shikimate pH 7.5, 22°C, mutant S336A Arabidopsis thaliana
399
-
NADP+ pH 7.5, 22°C, wild-type DELTA88DHQ-SDH enzyme variant Arabidopsis thaliana
428
-
shikimate pH 7.5, 22°C, wild-type DELTA88DHQ-SDH enzyme variant Arabidopsis thaliana

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Arabidopsis thaliana

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Arabidopsis thaliana
NADPH
-
Arabidopsis thaliana