Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21-CodonPlus (DE3)-RIL cells | Thermus thermophilus |
Crystallization (Comment) | Organism |
---|---|
in complex with amino group carrier protein LysW-gamma-alpha-aminoadipate plus NADP+ and amino group carrier protein LysW-gamma-aminoadipic semialdehyde plus NADP+, hanging drop vapor diffusion method, using 20% (w/v) PEG 2000 monomethyl ester and 200 mM calcium acetate | Thermus thermophilus |
Protein Variants | Comment | Organism |
---|---|---|
C148A | inactive | Thermus thermophilus |
H209A | the mutant exhibits similar activity to that of the wildtype enzyme | Thermus thermophilus |
K271A | the mutation results in a 4.6fold decrease in kcat and a 70fold increase in Km value compared to the wild type enzyme | Thermus thermophilus |
R195A | the mutation results in a 5fold decrease in kcat and a 40fold increase in Km value compared to the wild type enzyme | Thermus thermophilus |
R258A | the mutation results in a slight decrease in kcat and a 17fold increase in Km value compared to the wild type enzyme | Thermus thermophilus |
R278A | inactive | Thermus thermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0078 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme H209A, at pH 7.5 and 60°C | Thermus thermophilus | |
0.0098 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | wild type enzyme, at pH 9.5 and 60°C | Thermus thermophilus | |
0.011 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme H209A, at pH 9.5 and 60°C | Thermus thermophilus | |
0.011 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | wild type enzyme, at pH 7.5 and 60°C | Thermus thermophilus | |
0.014 | - |
NADP+ | wild type enzyme, at pH 9.5 and 60°C | Thermus thermophilus | |
0.037 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme R102A, at pH 9.5 and 60°C | Thermus thermophilus | |
0.17 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme R258A, at pH 9.5 and 60°C | Thermus thermophilus | |
0.39 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme R195A, at pH 9.5 and 60°C | Thermus thermophilus | |
0.69 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme K271A, at pH 9.5 and 60°C | Thermus thermophilus | |
12 | - |
phosphate | wild type enzyme, at pH 9.5 and 60°C | Thermus thermophilus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
[amino group carrier protein LysW]-gamma-alpha-aminoadipate + NADPH + H+ | Thermus thermophilus | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde + phosphate + NADP+ | - |
r | |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde + phosphate + NADP+ | Thermus thermophilus | - |
[amino group carrier protein LysW]-gamma-alpha-aminoadipate phosphate + NADPH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermus thermophilus | O50146 | - |
- |
Purification (Comment) | Organism |
---|---|
Blue-Sepharose column chromatography and Superdex S200 gel filtration | Thermus thermophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
[amino group carrier protein LysW]-gamma-alpha-aminoadipate + NADPH + H+ | - |
Thermus thermophilus | [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde + phosphate + NADP+ | - |
r | |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde + phosphate + NADP+ | - |
Thermus thermophilus | [amino group carrier protein LysW]-gamma-alpha-aminoadipate phosphate + NADPH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
homotetramer | 4 * 38000, SDS-PAGE | Thermus thermophilus |
Synonyms | Comment | Organism |
---|---|---|
lysY | - |
Thermus thermophilus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0063 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme R102A, at pH 9.5 and 60°C | Thermus thermophilus | |
0.0082 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme H209A, at pH 7.5 and 60°C | Thermus thermophilus | |
0.011 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | wild type enzyme, at pH 7.5 and 60°C | Thermus thermophilus | |
0.19 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme R195A, at pH 9.5 and 60°C | Thermus thermophilus | |
0.21 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme K271A, at pH 9.5 and 60°C | Thermus thermophilus | |
0.5 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme H209A, at pH 9.5 and 60°C | Thermus thermophilus | |
0.66 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme R258A, at pH 9.5 and 60°C | Thermus thermophilus | |
0.96 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | wild type enzyme, at pH 9.5 and 60°C | Thermus thermophilus | |
1.1 | - |
phosphate | wild type enzyme, at pH 9.5 and 60°C | Thermus thermophilus | |
1.2 | - |
NADP+ | wild type enzyme, at pH 9.5 and 60°C | Thermus thermophilus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Thermus thermophilus | |
NADPH | - |
Thermus thermophilus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0089 | - |
phosphate | wild type enzyme, at pH 9.5 and 60°C | Thermus thermophilus | |
0.17 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme R102A, at pH 9.5 and 60°C | Thermus thermophilus | |
0.31 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme K271A, at pH 9.5 and 60°C | Thermus thermophilus | |
0.49 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme R195A, at pH 9.5 and 60°C | Thermus thermophilus | |
1.1 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme H209A, at pH 7.5 and 60°C | Thermus thermophilus | |
4 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme R258A, at pH 9.5 and 60°C | Thermus thermophilus | |
15 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | wild type enzyme, at pH 7.5 and 60°C | Thermus thermophilus | |
47 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | mutant enzyme H209A, at pH 9.5 and 60°C | Thermus thermophilus | |
81 | - |
NADP+ | wild type enzyme, at pH 9.5 and 60°C | Thermus thermophilus | |
98 | - |
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde | wild type enzyme, at pH 9.5 and 60°C | Thermus thermophilus |