Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | the presence of an aldehyde substrate and NAD+ promotes isomerization of the enzyme into the active conformation | Homo sapiens |
Cloned (Comment) | Organism |
---|---|
recombinant expression of SUMO-His6-tagged ALDH9A1 in Escherichia coli strain BL21(DE3) | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant detagged enzyme in complex with NAD+ and inhibitor diethylaminobenzaldehyde, hanging drop vapour diffusion method, mixing of 6 mg/ml protein in 50 mM Tris-HCl, pH 8.0, 600 mM NaCl, 5% glycerol, 0.5 mM TCE, 5 mM DEAB, and 10 mM NAD+, with reservoir solution containing 0.1 M NaCl, 0.05 M Bis-Tris, pH 6.5, 0.1 M ammonium acetate, 0.05 M HEPES, pH 7.5, and 25% w/v PEG 3350, method optimization, X-ray diffraction structure determination and analysis at 2.50-2.64 A resolution, molecular replacement using structures of cod liver betaine ALDH (PDB ID 1A4S) and apo-ALDH9A1 (PDB ID 6QAP) as search models, modeling | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
diethylaminobenzaldehyde | DEAB, the broad-spectrum ALDH inhibitor reversibly inhibits ALDH9A1 in a time-dependent manner by a covalent reversible mechanism of inhibition, mechanism analysis, overview | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state Michaelis-Menten kinetics | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Homo sapiens | 5829 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
223000 | - |
recombinant detagged enzyme, gel filtration | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
3,4-dihydroxyphenylacetaldehyde + NAD+ + H2O | Homo sapiens | a dopamine metabolite | 3,4-dihydroxyphenylacetate + NADH + H+ | - |
? | |
4-trimethylaminobutyraldehyde + NAD+ + H2O | Homo sapiens | a carnitine precursor | 4-N-trimethylaminobutyrate + NADH + H+ | - |
? | |
aminobutyraldehyde + NAD+ + H2O | Homo sapiens | a GABA precursor | aminobutyrate + NADH + H+ | - |
? | |
betaine aldehyde + NAD+ + H2O | Homo sapiens | - |
betaine + NADH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P49189 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant SUMO-His6-tagged ALDH9A1 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, cleavage of the SUMO-His tag, dialysis, ultrafiltration, and gel filtration | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3,4-dihydroxyphenylacetaldehyde + NAD+ + H2O | - |
Homo sapiens | 3,4-dihydroxyphenylacetate + NADH + H+ | - |
? | |
3,4-dihydroxyphenylacetaldehyde + NAD+ + H2O | a dopamine metabolite | Homo sapiens | 3,4-dihydroxyphenylacetate + NADH + H+ | - |
? | |
4-trimethylaminobutyraldehyde + NAD+ + H2O | strongly preferred substrate | Homo sapiens | 4-N-trimethylaminobutyrate + NADH + H+ | - |
? | |
4-trimethylaminobutyraldehyde + NAD+ + H2O | a carnitine precursor | Homo sapiens | 4-N-trimethylaminobutyrate + NADH + H+ | - |
? | |
aminobutyraldehyde + NAD+ + H2O | - |
Homo sapiens | aminobutyrate + NADH + H+ | - |
? | |
aminobutyraldehyde + NAD+ + H2O | a GABA precursor | Homo sapiens | aminobutyrate + NADH + H+ | - |
? | |
betaine aldehyde + NAD+ + H2O | - |
Homo sapiens | betaine + NADH + H+ | - |
? | |
hexanal + NAD+ + H2O | - |
Homo sapiens | hexanoate + NADH + H+ | - |
? | |
additional information | diethylaminobenzaldehyde (DEAB) is not a substrate for ALDH9A1 | Homo sapiens | ? | - |
- |
Subunits | Comment | Organism |
---|---|---|
More | conformation of the inter-domain linker in the P1 ALDH9A1-NAD+ structure, modeling, overview. The in-solution quaternary structure of ALDH9A1 is determined using SAXS | Homo sapiens |
tetramer | enzyme ALDH9A1 forms the classic ALDH superfamily dimer-of-dimers tetramer in solution. Analytical ultracentrifugation, small-angle X-ray scattering (SAXS), and negative stain electron microscopy are used for analysis | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
aldehyde dehydrogenase 9A1 | - |
Homo sapiens |
Aldh9a1 | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
27 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | enzyme binding structure, enzyme-NAD+ crystal structure analysis, overview. NAD+ binds in the expected site at the C-termini of the beta-strands of the Rossmann fold. NAD+ forms several electrostatic interactions with the protein. The adenine ribose hydrogen bonds with Lys180. The diphosphate interacts with Trp156, Ser233, and Thr236. The nicotinamide ribose of the one complete NAD+ forms a hydrogen bond with Glu391, a residue identically conserved in the ALDH superfamily | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
evolution | aldehyde dehydrogenase 9A1 (ALDH9A1) belongs to the aldehyde dehydrogenase (ALDH) structural superfamily, which is a large group of enzymes that catalyze the NAD+-dependent oxidation of aldehydes to carboxylic acids. The superfamily comprises hundreds of distinct genes, including 19 ALDHs expressed in humans. ALDHs share a common protein fold and catalytic mechanism, but subtle differences in their active sites result in different preferences for the aldehyde substrate. Although ALDH9A1 exhibits the basic ALDH superfamily fold, the structure reveals two remarkable features. First, the final alpha-helix and beta-strand of the Rossmann dinucleotide-binding fold are disordered. Referred to as alphaE-betaE in the closely-related betaine ALDH, these secondary structural elements form extensive interactions with NAD+ in other ALDHs | Homo sapiens |
additional information | the active conformation of the enzyme, in which the Rossmann dinucleotide-binding domain is fully ordered and the inter-domain linker adopts the canonical beta-hairpin observed in other ALDH structures. The presence of an aldehyde substrate and NAD+ promotes isomerization of the enzyme into the active conformation | Homo sapiens |
physiological function | the major in vivo function of this enzyme is to catalyze the penultimate step of carnitine biosynthesis, the oxidation of TMBAL to 4-N-trimethylaminobutyrate. Carnitine functions in the transport of long-chain fatty acids from the cytosol to the mitochondrial matrix for the synthesis of acyl-CoAs for beta-oxidation. ALDH9A1 functions indirectly in beta-oxidation | Homo sapiens |