Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Arabidopsis thaliana |
Crystallization (Comment) | Organism |
---|---|
in complex with shikimate, and in ternary complex with shikimate and NADP+ | Arabidopsis thaliana |
Protein Variants | Comment | Organism |
---|---|---|
H335A | site-directed mutagenesis, tenfold decrease in kcat value | Arabidopsis thaliana |
N406A | site-directed mutagenesis, very strong decrease in kcat value | Arabidopsis thaliana |
Q578L | site-directed mutagenesis, sixtyfold decrease in kcat value | Arabidopsis thaliana |
Q582L | site-directed mutagenesis, very strong decrease in kcat value | Arabidopsis thaliana |
T381A | site-directed mutagenesis, more than twentyfold decrease in kcat value | Arabidopsis thaliana |
T381S | site-directed mutagenesis, fourfold decrease in kcat value, slight decrease in Km value | Arabidopsis thaliana |
T407A | site-directed mutagenesis, 6.5fold decrease in kcat value, increase in Km-value | Arabidopsis thaliana |
T422S | site-directed mutagenesis, tenfold decrease in kcat value | Arabidopsis thaliana |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.062 | - |
NADP+ | mutant T381S, 22°C | Arabidopsis thaliana | |
0.068 | - |
NADP+ | mutant H335A, 22°C | Arabidopsis thaliana | |
0.0928 | - |
NADP+ | mutant T381A, 22°C | Arabidopsis thaliana | |
0.107 | - |
NADP+ | mutant T422S, 22°C | Arabidopsis thaliana | |
0.131 | - |
NADP+ | wild-type, 22°C | Arabidopsis thaliana | |
0.156 | - |
NADP+ | mutant T407A, 22°C | Arabidopsis thaliana | |
0.159 | - |
NADP+ | mutant Q78L, 22°C | Arabidopsis thaliana | |
0.459 | - |
shikimate | mutant T381S, 22°C | Arabidopsis thaliana | |
0.556 | - |
shikimate | mutant T381A, 22°C | Arabidopsis thaliana | |
0.626 | - |
shikimate | mutant H335A, 22°C | Arabidopsis thaliana | |
0.685 | - |
shikimate | wild-type, 22°C | Arabidopsis thaliana | |
0.92 | - |
shikimate | mutant Q78L, 22°C | Arabidopsis thaliana | |
1.022 | - |
shikimate | mutant T407A, 22°C | Arabidopsis thaliana | |
1.09 | - |
shikimate | mutant T422S, 22°C | Arabidopsis thaliana | |
4.466 | - |
shikimate | mutant Q582L, 22°C | Arabidopsis thaliana | |
7.12 | - |
shikimate | mutant N406A, 22°C | Arabidopsis thaliana |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Arabidopsis thaliana | - |
- |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
shikimate + NADP+ = 3-dehydroshikimate + NADPH + H+ | the pro-R hydrogen of the nicotinamide C4 is 3.35 A from the C3 of shikimate. The catalytic K385 and D423 residues are proximal to the C3-hydroxyl of shikimate, which is deprotonated in the oxidation reaction. The enzyme adopts a concave architecture that places the active sites in a face-to-face arrangement | Arabidopsis thaliana |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
shikimate + NADP+ | - |
Arabidopsis thaliana | 3-dehydroshikimate + NADPH + H+ | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.174 | - |
shikimate | mutant N406A, 22°C | Arabidopsis thaliana | |
0.346 | - |
shikimate | mutant Q582L, 22°C | Arabidopsis thaliana | |
4.51 | - |
NADP+ | mutant Q78L, 22°C | Arabidopsis thaliana | |
7.3 | - |
shikimate | mutant Q78L, 22°C | Arabidopsis thaliana | |
15.1 | - |
NADP+ | mutant T381A, 22°C | Arabidopsis thaliana | |
16.8 | - |
shikimate | mutant T381A, 22°C | Arabidopsis thaliana | |
34 | - |
NADP+ | mutant H335A, 22°C | Arabidopsis thaliana | |
42 | - |
shikimate | mutant H335A, 22°C | Arabidopsis thaliana | |
43.3 | - |
NADP+ | mutant T422S, 22°C | Arabidopsis thaliana | |
46.3 | - |
shikimate | mutant T422S, 22°C | Arabidopsis thaliana | |
52.1 | - |
NADP+ | mutant T407A, 22°C | Arabidopsis thaliana | |
66.6 | - |
shikimate | mutant T407A, 22°C | Arabidopsis thaliana | |
105 | - |
NADP+ | mutant T381S, 22°C | Arabidopsis thaliana | |
114 | - |
shikimate | mutant T381S, 22°C | Arabidopsis thaliana | |
399 | - |
NADP+ | wild-type, 22°C | Arabidopsis thaliana | |
428 | - |
shikimate | wild-type, 22°C | Arabidopsis thaliana |