Literature summary for 1.2.1.47 extracted from

Koncitikova, R.; Vigouroux, A.; Kopecna, M.; Sebela, M.; Morera, S.; Kopecny, D.
Kinetic and structural analysis of human ALDH9A1 (2019), Biosci. Rep., 39, BSR20190558 .

Cloned(Commentary)

Cloned (Comment)	Organism
gene ALDH9A1, recombinant expression of His-tagged enzyme in Escherichia coli strain Rosetta2 (DE3) pLysS	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified human ALDH9A1 in apoform and in complex with NAD+ or 4-(trimethylamino)butyraldehyde, hanging drop vapour diffusion method, mixing of 28 mg/ml protein in 50 mM Tris/HCl, pH 7.5, and 150 mM NaCl, with 50 mM NAD+ and an equal volume of a precipitant solution containing 12% w/v PEG 4000, 0.1 M sodium citrate, pH 5.6, and 2.5% isopropanol, X-ray diffraction structure determination and analysis at 2.3 A, 2.9 A, and 2.5 A resolution, molecular replacement using the structure of BADH from Gadus morhua subsp. callarias liver as a search model (PDB IDs 1BPW and 1A4S), structure modeling	Homo sapiens

Crystallization (Comment)

Organism

purified human ALDH9A1 in apoform and in complex with NAD+ or 4-(trimethylamino)butyraldehyde, hanging drop vapour diffusion method, mixing of 28 mg/ml protein in 50 mM Tris/HCl, pH 7.5, and 150 mM NaCl, with 50 mM NAD+ and an equal volume of a precipitant solution containing 12% w/v PEG 4000, 0.1 M sodium citrate, pH 5.6, and 2.5% isopropanol, X-ray diffraction structure determination and analysis at 2.3 A, 2.9 A, and 2.5 A resolution, molecular replacement using the structure of BADH from Gadus morhua subsp. callarias liver as a search model (PDB IDs 1BPW and 1A4S), structure modeling

Homo sapiens

General Stability

General Stability	Organism
the enzyme is very sensitive and becomes nearly inactive after two re-freezing cycles. The highest melting temperatures are observed in buffers at pH 7.0 and 7.5	Homo sapiens

Inhibitors

Inhibitors	Comment	Organism
4-(dimethylamino)butyraldehyde	-	Homo sapiens
4-(trimethylamino)butyraldehyde	-	Homo sapiens
4-guanidinobutyraldehyde	-	Homo sapiens
additional information	the enzyme shows substrate inhibition	Homo sapiens
N,N,N-trimethyl-3-aminopropionaldehyde	-	Homo sapiens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
additional information	-	additional information	Michaelis-Menten kinetics	Homo sapiens
0.006	-	4-(trimethylamino)butyraldehyde	pH 7.5, temperature not specified in the publication	Homo sapiens
0.011	-	3,4-dihydroxyphenylacetaldehyde	pH 7.5, temperature not specified in the publication	Homo sapiens
0.017	-	acetaldehyde	pH 7.5, temperature not specified in the publication	Homo sapiens
0.021	-	4-guanidinobutyraldehyde	pH 7.5, temperature not specified in the publication	Homo sapiens
0.021	-	4-(dimethylamino)butyraldehyde	pH 7.5, temperature not specified in the publication	Homo sapiens
0.021	-	N,N-dimethyl-3-aminopropionaldehyde	pH 7.5, temperature not specified in the publication	Homo sapiens
0.026	-	2-hexenal	pH 7.5, temperature not specified in the publication	Homo sapiens
0.032	-	NAD+	pH 7.5, temperature not specified in the publication	Homo sapiens
0.035	-	Valeraldehyde	pH 7.5, temperature not specified in the publication	Homo sapiens
0.05	-	hexanal	pH 7.5, temperature not specified in the publication	Homo sapiens
0.053	-	N,N,N-trimethyl-3-aminopropionaldehyde	pH 7.5, temperature not specified in the publication	Homo sapiens
0.056	-	3-aminopropionaldehyde	pH 7.5, temperature not specified in the publication	Homo sapiens
0.067	-	4-Aminobutyraldehyde	pH 7.5, temperature not specified in the publication	Homo sapiens
0.216	-	Butyraldehyde	pH 7.5, temperature not specified in the publication	Homo sapiens

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
cytosol	-	Homo sapiens	5829	-

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
214000	-	recombinant His-tagged enzyme, gel filtration	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
4-(trimethylamino)butyraldehyde + NAD+ + H2O	Homo sapiens	preferred substrate	4-(trimethylamino)butyrate + NADH + 2 H+	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P49189	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli strain Rosetta2 (DE3) pLysS by cobalt affinity and nickel affinity chromatography, followed by ultrafiltration and gel filtration	Homo sapiens

Source Tissue

Source Tissue	Comment	Organism	Textmining
heart	-	Homo sapiens	-
kidney	-	Homo sapiens	-
liver	-	Homo sapiens	-
pancreas	-	Homo sapiens	-
skeletal muscle	-	Homo sapiens	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
2-hexenal + NAD+ + H2O	-	Homo sapiens	2-hexenoate + NADH + H+	-	?
3,4-dihydroxyphenylacetaldehyde + NAD+ + H2O	-	Homo sapiens	3,4-dihydroxyphenylacetate + NADH + H+	-	?
3-aminopropionaldehyde + NAD+ + H2O	-	Homo sapiens	3-aminopropionate + NADH + H+	-	?
3-guanidinopropionaldehyde + NAD+ + H2O	-	Homo sapiens	3-guanidinopropionate + NADH + H+	-	?
4-(dimethylamino)butyraldehyde + NAD+ + H2O	-	Homo sapiens	4-(dimethylamino)butyrate + NADH + 2 H+	-	?
4-(trimethylamino)butyraldehyde + NAD+ + H2O	preferred substrate	Homo sapiens	4-(trimethylamino)butyrate + NADH + 2 H+	-	?
4-amino-2-hydroxybutyraldehyde + NAD+ + H2O	-	Homo sapiens	4-amino-2-hydroxybutyrate + NADH + H+	-	?
4-aminobutyraldehyde + NAD+ + H2O	-	Homo sapiens	4-aminobutyrate + NADH + H+	-	?
4-guanidinobutyraldehyde + NAD+ + H2O	-	Homo sapiens	4-guanidinobutyrate + NADH + H+	-	?
acetaldehyde + NAD+ + H2O	-	Homo sapiens	acetate + NADH + H+	-	?
butyraldehyde + NAD+ + H2O	-	Homo sapiens	butyrate + NADH + H+	-	?
hexanal + NAD+ + H2O	-	Homo sapiens	hexanoate + NADH + H+	-	?
additional information	ALDH9A1 exhibits wide substrate specificity to aminoaldehydes, aliphatic and aromatic aldehydes with a clear preference for gamma-trimethylaminobutyraldehyde (TMABAL). The enzyme is multifunctional also catalyzing the reactions of EC 1.2.1.3 and EC 1.2.1.19. Substrate specificity, overview	Homo sapiens	?	-	-
N,N,N-trimethyl-3-aminopropionaldehyde + NAD+ + H2O	-	Homo sapiens	N,N,N-trimethyl-3-aminopropionate + NADH + H+	-	?
N,N-dimethyl-3-aminopropionaldehyde + NAD+ + H2O	-	Homo sapiens	N,N-dimethyl-3-aminopropionate + NADH + H+	-	?
valeraldehyde + NAD+ + H2O	-	Homo sapiens	valerate + NADH + H+	-	?

Subunits

Subunits	Comment	Organism
More	each ALDH monomer displays a typical ALDHs fold composed of an oligomerization domain (residues 128-145 and 479-494), a coenzyme domain (residues 1-127, 146-257, 470-478), a catalytic domain (residues 258-448) with the catalytic Cys288, and an interdomain linker highly conserved in amino-acid sequence and folding. Nonetheless, structural comparison reveals a position and a unique fold of the interdomain linker of ALDH9A1. The oligomerization domain wraps over the groove between the catalytic and coenzyme domains of the other monomer forming the dimer	Homo sapiens
tetramer	4 * 56000, dimer-of-dimers, recombinant His-tagged enzyme, SDS-PAGE	Homo sapiens

Synonyms

Synonyms	Comment	Organism
Aldh9a1	-	Homo sapiens
gamma-trimethylaminobutyraldehyde dehydrogenase	-	Homo sapiens
TMABAL dehydrogenase	-	Homo sapiens
TMABALDH	-	Homo sapiens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	-	Homo sapiens

Cofactor

Cofactor	Comment	Organism	Structure
NAD+	-	Homo sapiens

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism
additional information	-	additional information	inhibition kinetics	Homo sapiens
0.021	-	4-guanidinobutyraldehyde	pH 7.5, temperature not specified in the publication	Homo sapiens
0.47	-	N,N,N-trimethyl-3-aminopropionaldehyde	pH 7.5, temperature not specified in the publication	Homo sapiens
7.23	-	4-(trimethylamino)butyraldehyde	pH 7.5, temperature not specified in the publication	Homo sapiens
10.88	-	4-(dimethylamino)butyraldehyde	pH 7.5, temperature not specified in the publication	Homo sapiens

Expression

Organism	Comment	Expression
Homo sapiens	PPARalpha regulates the expression of ALDH9A1 gene	additional information

General Information

General Information	Comment	Organism
metabolism	the enzyme is involved in the carnitine synthesis pathway, it is abundantly expressed in tissues showing high rates of beta-oxidation such as liver and kidney	Homo sapiens
additional information	structure analysis, overview. Structural comparison reveals a position and a unique fold of the interdomain linker of ALDH9A1. This unique difference is not compatible with the presence of a bound substrate and a large conformational rearrangement of the linker up to 30 A has to occur to allow the access of the substrate channel. Moreover, the alphabetaE region consisting of an alpha-helix and a beta-strand of the coenzyme domain at the dimer interface are disordered, likely due to the loss of interactions with the inter-domain linker, which leads to incomplete beta-nicotinamide adenine dinucleotide (NAD+) binding pocket	Homo sapiens