Cloned (Comment) | Organism |
---|---|
expression of C-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Arabidopsis thaliana |
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 | 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 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Arabidopsis thaliana | - |
bifunctional enzyme | - |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
shikimate + NADP+ = 3-dehydroshikimate + NADPH + H+ | active site structure, reaction and kinetic mechanisms | Arabidopsis thaliana |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Arabidopsis thaliana |
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 | Comment | Organism |
---|---|---|
dehydroquinate dehydratase-shikimate dehydrogenase | - |
Arabidopsis thaliana |
DHQ-SDH | - |
Arabidopsis thaliana |
SDH | - |
Arabidopsis thaliana |
shikimate dehydrogenase | - |
Arabidopsis thaliana |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at | Arabidopsis thaliana |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Arabidopsis thaliana |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Arabidopsis thaliana | |
NADPH | - |
Arabidopsis thaliana |