Cloned (Comment) | Organism |
---|---|
overexpressed in Escherichia coli | Pseudomonas aeruginosa |
Crystallization (Comment) | Organism |
---|---|
two crystal structures of the enzyme are solved at 2.2-2.3 A resolution and reveal an N-terminal Rossmann fold domain connected by a long alpha-helix to the C-terminal all-alpha-domain. The apostructure shows the presence of additional density modeled as HEPES bound in the interdomain cleft close to the predicted catalytic Lys171, revealing the molecular details of the enzyme substrate-binding site. The structure of the enzyme-NAD complex demonstrates that the opposite side of the enzyme active site accommodates the cofactor, which is also bound near Lys171. Crystals of the enzyme are grown at 21C by the hanging drop vapor diffusion method with 0.002 ml of protein sample mixed with an equal volume of the reservoir buffer. The crystals of the wild-type enzyme grew after 1 week in the presence of 4 M ammonium acetate and 0.1 M sodium acetate (pH 5.4). The crystals of the complex of the enzyme with NAD+ are obtained by soaking the crystals in 10 mM NAD+. For diffraction studies, the crystals are stabilized with the crystallization buffer supplemented with 12% ethylene glycol as a cryoprotectant and flash frozen in liquid nitrogen | Pseudomonas aeruginosa |
Protein Variants | Comment | Organism |
---|---|---|
D247A | inactive protein | Pseudomonas aeruginosa |
K171A | inactive protein | Pseudomonas aeruginosa |
K246A | inactive protein | Pseudomonas aeruginosa |
N175A | mutant enzyme with very low activity | Pseudomonas aeruginosa |
S122A | mutant enzyme with very low activity | Pseudomonas aeruginosa |
T96A | mutant enzyme with very low activity | Pseudomonas aeruginosa |
W214A | inactive protein | Pseudomonas aeruginosa |
Y219A | mutant enzyme with very low activity | Pseudomonas aeruginosa |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.4 | - |
DL-threonine | pH 11.0, 37°C, wild-type enzyme | Pseudomonas aeruginosa | |
2.5 | - |
L-serine | pH 11.0, 37°C, wild-type enzyme | Pseudomonas aeruginosa | |
3.4 | - |
NAD+ | pH 11.0, 37°C, wild-type enzyme | Pseudomonas aeruginosa | |
4.2 | - |
L-serine | pH 11.0, 37°C, mutant enzyme N175A | Pseudomonas aeruginosa | |
10.6 | - |
L-serine | pH 11.0, 37°C, mutant enzyme S122A | Pseudomonas aeruginosa | |
10.8 | - |
D-glycerate | pH 11.0, 37°C, wild-type enzyme | Pseudomonas aeruginosa | |
11 | - |
L-serine | pH 11.0, 37°C, mutant enzyme Y219A | Pseudomonas aeruginosa | |
12.3 | - |
L-serine | pH 11.0, 37°C, mutant enzyme T96A | Pseudomonas aeruginosa | |
17.4 | - |
methyl 2,2-dimethyl-3-hydroxypropionate | pH 11.0, 37°C, wild-type enzyme | Pseudomonas aeruginosa |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-serine + NAD+ | Pseudomonas aeruginosa | the enzyme might be involved in serine/threonine degradation. Since growth experiments with various nitrogen and carbon sources (including L-serine) reveal no difference between the Pseudomonas aeruginosa wild-type and PA0743 deletion strains, it is suggested hat this organism might contain other (complementing) serine dehydrogenases | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas aeruginosa | Q9I5I6 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Pseudomonas aeruginosa |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
2-aminomalonate semialdehyde = 2-aminoacetaldehyde + CO2 | (1b), spontaneous | Pseudomonas aeruginosa | |
L-serine + NAD+ = 2-aminoacetaldehyde + CO2 + NADH + H+ | overall reaction | Pseudomonas aeruginosa | |
L-serine + NAD+ = 2-aminomalonate semialdehyde + NADH + H+ | (1a) | Pseudomonas aeruginosa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-glycerate + NAD+ | - |
Pseudomonas aeruginosa | ? | - |
? | |
DL-threonine + NAD+ | - |
Pseudomonas aeruginosa | ? | - |
? | |
L-serine + NAD+ | the enzyme might be involved in serine/threonine degradation. Since growth experiments with various nitrogen and carbon sources (including L-serine) reveal no difference between the Pseudomonas aeruginosa wild-type and PA0743 deletion strains, it is suggested hat this organism might contain other (complementing) serine dehydrogenases | Pseudomonas aeruginosa | ? | - |
? | |
L-serine + NAD+ | the enzyme might be involved in serine/methylserine degradation. Since growth experiments with various nitrogen and carbon sources (including L-serine) reveal no difference between the Pseudomonas aeruginosa wild-type and PA0743 deletion strains, it is suggested hat this organism might contain other (complementing) serine dehydrogenases | Pseudomonas aeruginosa | ? | - |
? | |
methyl 2,2-dimethyl-3-hydroxypropionate + NAD+ | - |
Pseudomonas aeruginosa | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
NAD+-dependent L-serine dehydrogenase | - |
Pseudomonas aeruginosa |
PA0743 | - |
Pseudomonas aeruginosa |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Pseudomonas aeruginosa |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.4 | - |
L-serine | pH 11.0, 37°C, mutant enzyme S122A | Pseudomonas aeruginosa | |
0.8 | - |
L-serine | pH 11.0, 37°C, mutant enzyme Y219A | Pseudomonas aeruginosa | |
1.4 | - |
L-serine | pH 11.0, 37°C, mutant enzyme T96A | Pseudomonas aeruginosa | |
1.6 | - |
NAD+ | pH 11.0, 37°C, wild-type enzyme | Pseudomonas aeruginosa | |
2.7 | - |
L-serine | pH 11.0, 37°C, mutant enzyme N175A | Pseudomonas aeruginosa | |
5.8 | - |
D-glycerate | pH 11.0, 37°C, wild-type enzyme | Pseudomonas aeruginosa | |
9.6 | - |
DL-threonine | pH 11.0, 37°C, wild-type enzyme | Pseudomonas aeruginosa | |
10.4 | - |
L-serine | pH 11.0, 37°C, wild-type enzyme | Pseudomonas aeruginosa | |
11.6 | - |
methyl 2,2-dimethyl-3-hydroxypropionate | pH 11.0, 37°C, wild-type enzyme | Pseudomonas aeruginosa |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
10 | 11 | - |
Pseudomonas aeruginosa |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | the enzyme can also use NADP+ as a cofactor for the oxidation of L-serine, but this activity is significantly lower than that with NAD+ (46%) | Pseudomonas aeruginosa | |
NADP+ | the enzyme can also use NADP+ as a cofactor for the oxidation of L-serine, but this activity is significantly lower than that with NAD+ (46%) | Pseudomonas aeruginosa |
General Information | Comment | Organism |
---|---|---|
metabolism | the enzyme might be involved in serine/threonine degradation. Since growth experiments with various nitrogen and carbon sources (including L-serine) reveal no difference between the Pseudomonas aeruginosa wild-type and PA0743 deletion strains, it is suggested hat this organism might contain other (complementing) serine dehydrogenases | Pseudomonas aeruginosa |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.04 | - |
L-serine | pH 11.0, 37°C, mutant enzyme S122A | Pseudomonas aeruginosa | |
0.07 | - |
L-serine | pH 11.0, 37°C, mutant enzyme Y219A | Pseudomonas aeruginosa | |
0.1 | - |
L-serine | pH 11.0, 37°C, mutant enzyme T96A | Pseudomonas aeruginosa | |
0.3 | - |
D-glycerate | pH 11.0, 37°C, wild-type enzyme | Pseudomonas aeruginosa | |
0.5 | - |
NAD+ | pH 11.0, 37°C, wild-type enzyme | Pseudomonas aeruginosa | |
0.6 | - |
L-serine | pH 11.0, 37°C, mutant enzyme N175A | Pseudomonas aeruginosa | |
0.7 | - |
methyl 2,2-dimethyl-3-hydroxypropionate | pH 11.0, 37°C, wild-type enzyme | Pseudomonas aeruginosa | |
4 | - |
L-serine | pH 11.0, 37°C, wild-type enzyme | Pseudomonas aeruginosa | |
4 | - |
DL-threonine | pH 11.0, 37°C, wild-type enzyme | Pseudomonas aeruginosa |