Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli Rosetta cells | Sorghum bicolor |
Crystallization (Comment) | Organism |
---|---|
isoform CAD4, hanging drop vapor diffusion method, using 2% (v/v) tacsimate, pH 7.0, 5% (v/v) 2-propanol, 0.1 M imidazole, pH 7.0, and 8% (w/v) polyethylene glycol 3350 | Sorghum bicolor |
Protein Variants | Comment | Organism |
---|---|---|
A278V | the isoform CAD4 mutant shows strongly decreased catalytic efficiency compared to the wild type enzyme | Sorghum bicolor |
G301F | the isoform CAD4 mutant enzyme displays slightly elevated Km values for 4-coumaryl aldehyde but decreased Km for coniferyl aldehyde compared with those values of wild type. The mutant becomes more dedicated toward sinapyl aldehyde, with substantially decreased efficiency for coniferyl aldehyde compared to the wild type enzyme | Sorghum bicolor |
L119W | the isoform CAD4 mutant displays a slightly reduced Km value for both 4-coumaryl aldehyde and coniferyl aldehyde compared to the wild type enzyme. The mutant becomes more efficient toward coniferaldehyde due to the very small Km value, with substantially decreased efficiency for both sinapyl aldehyde and 4-coumaryl aldehyde | Sorghum bicolor |
L119W/G301F | the isoform CAD4 double mutant displays its substrate preference in the order coniferyl aldehyde over 4-coumaryl aldehyde over sinapyl aldehyde, with higher catalytic efficiency than that of wild type enzyme. The mutant displays 10 and 800fold increases in its catalytic efficiency for coniferyl aldehyde and 4-coumaryl aldehyde, respectively, and a 50% decrease in catalytic efficiency for sinapyl aldehyde as compared to the wild type enzyme | Sorghum bicolor |
W58L | the isoform CAD4 mutant enzyme displays slightly elevated Km values for 4-coumaryl aldehyde but decreased Km for coniferyl aldehyde compared with those values of wild type | Sorghum bicolor |
Y288P | the isoform CAD4 mutant becomes more dedicated toward sinapyl aldehyde, with substantially decreased efficiency for coniferyl aldehyde compared to the wild type enzyme | Sorghum bicolor |
Y95V | the isoform CAD4 mutant shows decreased catalytic efficiency compared to the wild type enzyme | Sorghum bicolor |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
4-coumaryl aldehyde | - |
Sorghum bicolor | |
coniferyl aldehyde | - |
Sorghum bicolor | |
sinapyl aldehyde | - |
Sorghum bicolor |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0078 | - |
sinapyl aldehyde | isoform CAD4 mutant Y288P, at pH 6.5 and 30°C | Sorghum bicolor | |
0.0104 | - |
coniferyl aldehyde | isoform CAD4 mutant L119W, at pH 6.5 and 30°C | Sorghum bicolor | |
0.0168 | - |
sinapyl aldehyde | isoform CAD4 mutant G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
0.0214 | - |
coniferyl aldehyde | isoform CAD4 mutant A278V, at pH 6.5 and 30°C | Sorghum bicolor | |
0.028 | - |
coniferyl aldehyde | isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
0.033 | - |
sinapyl aldehyde | wild type isoform CAD4, at pH 6.5 and 30°C | Sorghum bicolor | |
0.05976 | - |
sinapyl aldehyde | wild type isoform CAD2, at pH 6.5 and 30°C | Sorghum bicolor | |
0.068 | - |
coniferyl aldehyde | isoform CAD4 mutant Y95V, at pH 6.5 and 30°C | Sorghum bicolor | |
0.06975 | - |
4-coumaryl aldehyde | wild type isoform CAD2, at pH 6.5 and 30°C | Sorghum bicolor | |
0.0783 | - |
sinapyl aldehyde | isoform CAD4 mutant L119W, at pH 6.5 and 30°C | Sorghum bicolor | |
0.0793 | - |
sinapyl aldehyde | isoform CAD4 mutant A278V, at pH 6.5 and 30°C | Sorghum bicolor | |
0.0809 | - |
4-coumaryl aldehyde | isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
0.09518 | - |
coniferyl aldehyde | wild type isoform CAD2, at pH 6.5 and 30°C | Sorghum bicolor | |
0.105 | - |
sinapyl aldehyde | isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
0.112 | - |
sinapyl aldehyde | isoform CAD4 mutant Y95V, at pH 6.5 and 30°C | Sorghum bicolor | |
0.233 | - |
coniferyl aldehyde | wild type isoform CAD4, at pH 6.5 and 30°C | Sorghum bicolor | |
0.268 | - |
4-coumaryl aldehyde | isoform CAD4 mutant G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
0.387 | - |
coniferyl aldehyde | isoform CAD4 mutant Y288P, at pH 6.5 and 30°C | Sorghum bicolor | |
2.12 | - |
4-coumaryl aldehyde | isoform CAD4 mutant L119W, at pH 6.5 and 30°C | Sorghum bicolor | |
4.18 | - |
4-coumaryl aldehyde | isoform CAD4 mutant Y288P, at pH 6.5 and 30°C | Sorghum bicolor | |
4.63 | - |
4-coumaryl aldehyde | wild type isoform CAD4, at pH 6.5 and 30°C | Sorghum bicolor | |
5.52 | - |
sinapyl aldehyde | isoform CAD4 mutant W58L, at pH 6.5 and 30°C | Sorghum bicolor | |
5.68 | - |
coniferyl aldehyde | isoform CAD4 mutant G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
6.88 | - |
4-coumaryl aldehyde | isoform CAD4 mutant Y95V, at pH 6.5 and 30°C | Sorghum bicolor | |
7.1 | - |
4-coumaryl aldehyde | isoform CAD4 mutant W58L, at pH 6.5 and 30°C | Sorghum bicolor | |
7.97 | - |
4-coumaryl aldehyde | isoform CAD4 mutant A278V, at pH 6.5 and 30°C | Sorghum bicolor | |
9.17 | - |
coniferyl aldehyde | isoform CAD4 mutant W58L, at pH 6.5 and 30°C | Sorghum bicolor |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | dependent on | Sorghum bicolor |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Sorghum bicolor | A1ILL4 | - |
- |
Sorghum bicolor | C5XC49 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA agarose column chromatography and Mono Q column chromatography | Sorghum bicolor |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-coumaryl aldehyde + NADPH + H+ | - |
Sorghum bicolor | 4-coumaryl alcohol + NADP+ | - |
? | |
coniferyl aldehyde + NADPH + H+ | - |
Sorghum bicolor | coniferyl alcohol + NADP+ | ? | ? | |
sinapyl aldehyde + NADPH + H+ | - |
Sorghum bicolor | sinapyl alcohol + NADP+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CAD2 | isoform | Sorghum bicolor |
CAD4 | isoform | Sorghum bicolor |
cinnamyl alcohol dehydrogenase | - |
Sorghum bicolor |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.76 | - |
coniferyl aldehyde | isoform CAD4 mutant A278V, at pH 6.5 and 30°C | Sorghum bicolor | |
1.05 | - |
sinapyl aldehyde | isoform CAD4 mutant A278V, at pH 6.5 and 30°C | Sorghum bicolor | |
1.08 | - |
4-coumaryl aldehyde | isoform CAD4 mutant L119W, at pH 6.5 and 30°C | Sorghum bicolor | |
1.19 | - |
4-coumaryl aldehyde | isoform CAD4 mutant Y288P, at pH 6.5 and 30°C | Sorghum bicolor | |
1.59 | - |
sinapyl aldehyde | isoform CAD4 mutant Y288P, at pH 6.5 and 30°C | Sorghum bicolor | |
2.17 | - |
4-coumaryl aldehyde | wild type isoform CAD4, at pH 6.5 and 30°C | Sorghum bicolor | |
2.33 | - |
coniferyl aldehyde | isoform CAD4 mutant L119W, at pH 6.5 and 30°C | Sorghum bicolor | |
2.5 | - |
sinapyl aldehyde | isoform CAD4 mutant W58L, at pH 6.5 and 30°C | Sorghum bicolor | |
3.03 | - |
4-coumaryl aldehyde | isoform CAD4 mutant A278V, at pH 6.5 and 30°C | Sorghum bicolor | |
3.27 | - |
4-coumaryl aldehyde | isoform CAD4 mutant W58L, at pH 6.5 and 30°C | Sorghum bicolor | |
3.32 | - |
sinapyl aldehyde | isoform CAD4 mutant L119W, at pH 6.5 and 30°C | Sorghum bicolor | |
3.59 | - |
4-coumaryl aldehyde | isoform CAD4 mutant Y95V, at pH 6.5 and 30°C | Sorghum bicolor | |
3.68 | - |
coniferyl aldehyde | isoform CAD4 mutant W58L, at pH 6.5 and 30°C | Sorghum bicolor | |
3.76 | - |
coniferyl aldehyde | isoform CAD4 mutant Y288P, at pH 6.5 and 30°C | Sorghum bicolor | |
5.29 | - |
sinapyl aldehyde | isoform CAD4 mutant G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
9.26 | - |
coniferyl aldehyde | isoform CAD4 mutant Y95V, at pH 6.5 and 30°C | Sorghum bicolor | |
9.5 | - |
4-coumaryl aldehyde | isoform CAD4 mutant G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
11.93 | - |
4-coumaryl aldehyde | wild type isoform CAD2, at pH 6.5 and 30°C | Sorghum bicolor | |
12.54 | - |
sinapyl aldehyde | wild type isoform CAD4, at pH 6.5 and 30°C | Sorghum bicolor | |
12.77 | - |
coniferyl aldehyde | wild type isoform CAD2, at pH 6.5 and 30°C | Sorghum bicolor | |
18.98 | - |
sinapyl aldehyde | isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
20.28 | - |
coniferyl aldehyde | wild type isoform CAD4, at pH 6.5 and 30°C | Sorghum bicolor | |
25.83 | - |
coniferyl aldehyde | isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
30.16 | - |
4-coumaryl aldehyde | isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
31.75 | - |
sinapyl aldehyde | wild type isoform CAD2, at pH 6.5 and 30°C | Sorghum bicolor | |
39.97 | - |
sinapyl aldehyde | isoform CAD4 mutant Y95V, at pH 6.5 and 30°C | Sorghum bicolor | |
71.57 | - |
coniferyl aldehyde | isoform CAD4 mutant G301F, at pH 6.5 and 30°C | Sorghum bicolor |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | - |
Sorghum bicolor |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0121 | - |
coniferyl aldehyde | isoform CAD4 mutant Y95V, at pH 6.5 and 30°C | Sorghum bicolor | |
0.0162 | - |
coniferyl aldehyde | isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
0.0374 | - |
sinapyl aldehyde | isoform CAD4 mutant Y95V, at pH 6.5 and 30°C | Sorghum bicolor | |
0.083 | - |
sinapyl aldehyde | wild type isoform CAD2, at pH 6.5 and 30°C | Sorghum bicolor | |
0.099 | - |
4-coumaryl aldehyde | isoform CAD4 mutant G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
0.118 | - |
sinapyl aldehyde | wild type isoform CAD4, at pH 6.5 and 30°C | Sorghum bicolor | |
0.123 | - |
coniferyl aldehyde | wild type isoform CAD2, at pH 6.5 and 30°C | Sorghum bicolor | |
0.16 | - |
4-coumaryl aldehyde | isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
0.295 | - |
coniferyl aldehyde | isoform CAD4 mutant W58L, at pH 6.5 and 30°C | Sorghum bicolor | |
0.295 | - |
coniferyl aldehyde | wild type isoform CAD4, at pH 6.5 and 30°C | Sorghum bicolor | |
0.427 | - |
coniferyl aldehyde | isoform CAD4 mutant L119W, at pH 6.5 and 30°C | Sorghum bicolor | |
0.711 | - |
coniferyl aldehyde | isoform CAD4 mutant G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
1.056 | - |
sinapyl aldehyde | isoform CAD4 mutant L119W, at pH 6.5 and 30°C | Sorghum bicolor | |
1.23 | - |
sinapyl aldehyde | isoform CAD4 mutant G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
1.39 | - |
sinapyl aldehyde | isoform CAD4 mutant Y288P, at pH 6.5 and 30°C | Sorghum bicolor | |
1.455 | - |
sinapyl aldehyde | isoform CAD4 mutant W58L, at pH 6.5 and 30°C | Sorghum bicolor | |
2.91 | - |
sinapyl aldehyde | isoform CAD4 mutant A278V, at pH 6.5 and 30°C | Sorghum bicolor | |
4.17 | - |
coniferyl aldehyde | isoform CAD4 mutant A278V, at pH 6.5 and 30°C | Sorghum bicolor | |
6.03 | - |
coniferyl aldehyde | isoform CAD4 mutant Y288P, at pH 6.5 and 30°C | Sorghum bicolor | |
7.63 | - |
4-coumaryl aldehyde | isoform CAD4 mutant Y95V, at pH 6.5 and 30°C | Sorghum bicolor | |
10.02 | - |
4-coumaryl aldehyde | isoform CAD4 mutant W58L, at pH 6.5 and 30°C | Sorghum bicolor | |
20.59 | - |
4-coumaryl aldehyde | wild type isoform CAD4, at pH 6.5 and 30°C | Sorghum bicolor | |
31.92 | - |
4-coumaryl aldehyde | isoform CAD4 mutant A278V, at pH 6.5 and 30°C | Sorghum bicolor | |
32.67 | - |
4-coumaryl aldehyde | wild type isoform CAD2, at pH 6.5 and 30°C | Sorghum bicolor | |
68.08 | - |
sinapyl aldehyde | isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C | Sorghum bicolor |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.28 | - |
4-coumaryl aldehyde | isoform CAD4 mutant Y288P, at pH 6.5 and 30°C | Sorghum bicolor | |
0.38 | - |
4-coumaryl aldehyde | isoform CAD4 mutant A278V, at pH 6.5 and 30°C | Sorghum bicolor | |
0.4 | - |
coniferyl aldehyde | isoform CAD4 mutant W58L, at pH 6.5 and 30°C | Sorghum bicolor | |
0.45 | - |
sinapyl aldehyde | isoform CAD4 mutant W58L, at pH 6.5 and 30°C | Sorghum bicolor | |
0.46 | - |
4-coumaryl aldehyde | isoform CAD4 mutant W58L, at pH 6.5 and 30°C | Sorghum bicolor | |
0.47 | - |
4-coumaryl aldehyde | wild type isoform CAD4, at pH 6.5 and 30°C | Sorghum bicolor | |
0.51 | - |
4-coumaryl aldehyde | isoform CAD4 mutant L119W, at pH 6.5 and 30°C | Sorghum bicolor | |
0.52 | - |
4-coumaryl aldehyde | isoform CAD4 mutant Y95V, at pH 6.5 and 30°C | Sorghum bicolor | |
10 | - |
sinapyl aldehyde | isoform CAD4 mutant A278V, at pH 6.5 and 30°C | Sorghum bicolor | |
10 | - |
coniferyl aldehyde | isoform CAD4 mutant G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
10 | - |
coniferyl aldehyde | isoform CAD4 mutant Y288P, at pH 6.5 and 30°C | Sorghum bicolor | |
40 | - |
coniferyl aldehyde | isoform CAD4 mutant A278V, at pH 6.5 and 30°C | Sorghum bicolor | |
40 | - |
4-coumaryl aldehyde | isoform CAD4 mutant G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
40 | - |
sinapyl aldehyde | isoform CAD4 mutant L119W, at pH 6.5 and 30°C | Sorghum bicolor | |
90 | - |
coniferyl aldehyde | wild type isoform CAD4, at pH 6.5 and 30°C | Sorghum bicolor | |
130 | - |
coniferyl aldehyde | wild type isoform CAD2, at pH 6.5 and 30°C | Sorghum bicolor | |
140 | - |
coniferyl aldehyde | isoform CAD4 mutant Y95V, at pH 6.5 and 30°C | Sorghum bicolor | |
170 | - |
4-coumaryl aldehyde | wild type isoform CAD2, at pH 6.5 and 30°C | Sorghum bicolor | |
180 | - |
sinapyl aldehyde | isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
200 | - |
sinapyl aldehyde | isoform CAD4 mutant Y288P, at pH 6.5 and 30°C | Sorghum bicolor | |
220 | - |
coniferyl aldehyde | isoform CAD4 mutant L119W, at pH 6.5 and 30°C | Sorghum bicolor | |
310 | - |
sinapyl aldehyde | isoform CAD4 mutant G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
360 | - |
sinapyl aldehyde | isoform CAD4 mutant Y95V, at pH 6.5 and 30°C | Sorghum bicolor | |
370 | - |
4-coumaryl aldehyde | isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C | Sorghum bicolor | |
380 | - |
sinapyl aldehyde | wild type isoform CAD4, at pH 6.5 and 30°C | Sorghum bicolor | |
530 | - |
sinapyl aldehyde | wild type isoform CAD2, at pH 6.5 and 30°C | Sorghum bicolor | |
920 | - |
coniferyl aldehyde | isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C | Sorghum bicolor |