Cloned (Comment) | Organism |
---|---|
recombinant expressin of N-terminally His-tagged wild-type and mutant enzymes in Escherichia coli | Pseudomonas fluorescens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged E268A mutant enzyme in complex with NAD+ or NAD+ and 2-aminomuconate or 2-hydroxymuconate, hanging drop diffusion, mixing of 0.0015 ml of 40 mg/ml protein solution with 0.0015 ml of reservoir solution containing 20% PEG 3350 and 0.2 M sodium phosphate dibasic monohydrate, pH 9.1, at 18°C, 2-3 days, crystals are soaked in ligand solution for the formation of complex crystals, X-ray diffraction structure determination and analysis at 1.95-2.20 A resolution, molecular replacement using 5-carboxymethyl-2-hydroxymuconate semialdehyde dehydrogenase, PDB: 2D4E, as a search model, and molecular modelling | Pseudomonas fluorescens |
Protein Variants | Comment | Organism |
---|---|---|
E268A | site-directed mutagenesis, catalytically inactive mutant, formation of an enzyme-substrate adduct in the E268A mutant investigated by mass spectrometry | Pseudomonas fluorescens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten steady-state kinetics and kinetic analysis, overview | Pseudomonas fluorescens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-aminomuconate 6-semialdehyde + NAD+ + H2O | Pseudomonas fluorescens | - |
2-aminomuconate + NADH + 2 H+ | - |
? | |
2-aminomuconate 6-semialdehyde + NAD+ + H2O | Pseudomonas fluorescens KU-7 | - |
2-aminomuconate + NADH + 2 H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas fluorescens | Q83V33 | - |
- |
Pseudomonas fluorescens KU-7 | Q83V33 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His-tagged wild-type and mutant enzymes from Escherichia coli | Pseudomonas fluorescens |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
2-aminomuconate 6-semialdehyde + NAD+ + H2O = 2-aminomuconate + NADH + H+ | the catalytic cycle, overview | Pseudomonas fluorescens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-aminomuconate 6-semialdehyde + NAD+ + H2O | - |
Pseudomonas fluorescens | 2-aminomuconate + NADH + 2 H+ | - |
? | |
2-aminomuconate 6-semialdehyde + NAD+ + H2O | enzyme-substrate binding structure and catalytic mechanism, overview. Due to the unstable nature of its substrate 2-aminomuconate 6-semialdehyde, 2-AMS, the activity of AMSDH is detected using a coupled-enzyme assay that employs its upstream partner, alpha-amino beta-carboxymuconate epsilon-semialdehyde decarboxylase (ACMSD), to generate 2-AMS in situ | Pseudomonas fluorescens | 2-aminomuconate + NADH + 2 H+ | - |
? | |
2-aminomuconate 6-semialdehyde + NAD+ + H2O | - |
Pseudomonas fluorescens KU-7 | 2-aminomuconate + NADH + 2 H+ | - |
? | |
2-aminomuconate 6-semialdehyde + NAD+ + H2O | enzyme-substrate binding structure and catalytic mechanism, overview. Due to the unstable nature of its substrate 2-aminomuconate 6-semialdehyde, 2-AMS, the activity of AMSDH is detected using a coupled-enzyme assay that employs its upstream partner, alpha-amino beta-carboxymuconate epsilon-semialdehyde decarboxylase (ACMSD), to generate 2-AMS in situ | Pseudomonas fluorescens KU-7 | 2-aminomuconate + NADH + 2 H+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 56252, recombinant N-terminally His-tagged enzyme mutant E268A, mass spectrometry | Pseudomonas fluorescens |
More | analysis of crystal structures of a tetrahedral, thiohemiacetal intermediate, a thioacyl intermediate and an NAD+-bound complex from the active site mutant E268A. These covalent intermediates are characterized by single-crystal and solution-state electronic absorption spectroscopy. The crystal structures reveal that the substrate undergoes an E/Z isomerization at the enzyme active site before an sp3-to-sp2 transition during enzyme-mediated oxidation | Pseudomonas fluorescens |
Synonyms | Comment | Organism |
---|---|---|
2-aminomuconate-6-semialdehyde dehydrogenase | - |
Pseudomonas fluorescens |
AMSDH | - |
Pseudomonas fluorescens |
nbaE | gene name, UniProt | Pseudomonas fluorescens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
10 | - |
assay at | Pseudomonas fluorescens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Pseudomonas fluorescens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | enzyme-cofactor binding structure and catalytic mechanism, overview | Pseudomonas fluorescens |
General Information | Comment | Organism |
---|---|---|
metabolism | the enzyme is involved in the metabolic pathway for tryptophan degradation | Pseudomonas fluorescens |
additional information | analysis of catalytic mechanism, formation of catalytic intermediates, ligand binding, and active site structure, molecular modelling, overview | Pseudomonas fluorescens |
physiological function | several aldehydic intermediates in the metabolic pathway for tryptophan degradation can decay into neuroactive compounds that are associated with numerous neurological diseases. 2-Aminomuconate-6-semialdehyde dehydrogenase is involved in the pathway and is responsible for disarming the final aldehydic intermediate. The enzymeis responsible for oxidizing the unstable metabolic intermediate 2-aminomuconate-6-semialdehyde to 2-aminomuconate | Pseudomonas fluorescens |