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
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
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
DEAB, the broad-spectrum ALDH inhibitor reversibly inhibits ALDH9A1 in a time-dependent manner by a covalent reversible mechanism of inhibition, mechanism analysis, overview
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
enzyme overexpression causes the Kawasaki disease (KD), an acute vasculitis that preferentially affects coronary arteries. The disease is still the leading cause of acquired heart disease in children. Patients suffering Kawasaki disease show increased TMABADH enzyme protein levels compared to controls
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
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
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
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
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
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
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
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
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
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
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
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
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
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
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
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
recombinant His-tagged enzyme from Escherichia coli strain Rosetta2 (DE3) pLysS by cobalt affinity and nickel affinity chromatography, followed by ultrafiltration and gel filtration
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
detection of anti-TMABA-DH autoantibody is a potential strategy for a diagnosis of Kawasaki disease, usefulness of the anti-TMABA-DH antibody as a diagnostic marker
Neurotoxicity and metabolism of the catecholamine-derived 3,4-dihydroxyphenylacetaldehyde and 3,4-dihydroxyphenylglycolaldehyde: the role of aldehyde dehydrogenase