Active site of human liver aldehyde dehydrogenase

Abriola, Darryl P.; Fields, Robert; Stein, Stanley J.; MacKerell, Alexander D.; Pietruszko, Regina

doi:10.1021/bi00392a015

Cited by 96 publications

(58 citation statements)

References 22 publications

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“…3) shows that the residues Glu 270 , Cys 304 , and Gln 489 , which are thought to be involved in the catalytic role of ALDH enzymes (41)(42)(43), were highly conserved in Xenopus sequences. The region participating in T 3 binding by xCTBP/xALDH1 is located at amino acid positions 92-107.…”

Section: Discussionmentioning

confidence: 99%

Xenopus Cytosolic Thyroid Hormone-binding Protein (xCTBP) Is Aldehyde Dehydrogenase Catalyzing the Formation of Retinoic Acid

Yamauchi

Nakajima

Hayashi

et al. 1999

Journal of Biological Chemistry

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A major characteristic of 3,3Ј,5-triiodo-L-thyronine (T 3 ), 1 the active form of thyroid hormone at the cellular level, is the multiplicity of physiological processes. These include such diverse functions as postembryonic and fetal development and postnatal growth in mammals, amphibian metamorphosis, maturation of central nervous system, energy metabolism in homeotherms, and environmental adaptation in poikilotherms (1-4). It is now generally accepted that, at the molecular level, most of these actions of thyroid hormone are initiated by the interaction between thyroid hormone receptor and T 3 . Thyroid hormone receptor is a member of a multigene family of nuclear receptors that act as transcription factors in combination with transcriptional co-activators and co-repressors and chromatinmodifying factors (5-9). It is, however, not clear as to how T 3 enters the cell and reaches the nucleus and what determines the dynamics of cytoplasm-to-nucleus transfer of the hormone.

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Section: Discussionmentioning

confidence: 99%

Xenopus Cytosolic Thyroid Hormone-binding Protein (xCTBP) Is Aldehyde Dehydrogenase Catalyzing the Formation of Retinoic Acid

Yamauchi

Nakajima

Hayashi

et al. 1999

Journal of Biological Chemistry

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“…By performing site-directed mutagenesis with the rat liver enzyme, we now find that the residue is cysteine 302 (63), as originally suggested by Hempel and Pietruszko (25). Recently, however, it was found that in sheep liver cytosolic ALDH, serine 74 could be modified by a substrate (38 It was reported that glutamate 268 in the human cytosolic enzyme appeared to be essential for activity (1,2). This is also a highly conserved residue in all species.…”

Section: Discussionmentioning

confidence: 99%

Molecular cloning of the mitochondrial aldehyde dehydrogenase gene of Saccharomyces cerevisiae by genetic complementation

et al. 1991

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Mutants of Saccharomyces cerevisiae deficient in mitochondrial aldehyde dehydrogenase (ALDH) activity were isolated by chemical mutagenesis with ethyl methanesulfonate. The mutants were selected by their inability to grow on ethanol as the sole carbon source. The ALDH mutants were distinguished from alcohol dehydrogenase mutants by an aldehyde indicator plate test and by immunoscreening. The ALDH gene was isolated from a yeast genomic DNA library on a 5.7-kb insert of a recombinant DNA plasmid by functional complementation of the aldh mutation in S.cerevisiae. An open reading frame which specifies 533 codons was found within the 2.0-kb BamHI-BstEII fragment in the 5.7-kb genomic insert which can encode a protein with a molecular weight of 58,630. The N-terminal portion of the protein contains many positively charged residues which may serve as a signal sequence that targets the protein to the mitochondria. The amino acid sequence of the proposed mature yeast enzyme shows 30% identity to each of the known ALDH sequences from eukaryotes or prokaryotes. The amino acid residues corresponding to mammalian cysteine 302 and glutamates 268 and 487, implicated to be involved at the active site, were conserved. S. cerevisiae ALDH was found to be localized in the mitochondria as a tetrameric enzyme. Thius, that organelle is responsible for acetaldehyde oxidation, as was found in mammalian liver.Saccharomyces cerevisiae, unlike higher eukaryotes, can metabolize as well as produce ethanol. During fermentation, the consumption of sugars results in the accumulation of ethanol. When glucose or any other fermentable sugar is absent from the culture medium, S. cerevisiae can utilize ethanol as the carbon source aerobically. The metabolism of ethanol is well understood in mammals. In the liver, cytosolic alcohol dehydrogenase (ADH) oxidizes ethanol to acetaldehyde and then mitochondrial aldehyde dehydrogenase (ALDH) oxidizes the intermediate to acetate (14). In S. cerevisiae, it appears that a mitochondrial alcohol dehydrogenase (ADH3) is responsible for the initial oxidation (65,67). Little is known about the role of ALDHs in the next step.Various groups have purified and characterized yeast ALDH (12,49,54). It was reported to be a tetrameric enzyme with a subunit molecular mass of 60 kDa and a low Km for acetaldehyde. Unlike the liver enzymes, the yeast enzyme is activated by K+ ions (4, 32). Whereas the mammalian enzymes have been sequenced at the cDNA (16,19,29,34) as well as the protein (24, 33, 60) level, no sequence work has been reported for the yeast enzyme.One of our interests was to study the subcellular localization of acetaldehyde metabolism in S. cerevisiae. In the mammalian liver tissue, this was studied by selectively inhibiting the cytosolic or the mitochondrial isozyme of ALDH (11,52). An alternative approach would be to introduce the enzyme into a cell deficient in ALDH activity. S. cerevisiae would be a suitable model system, as it can metabolize ethanol and could serve as a host for the expression of foreign g...

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“…These residues were conserved in all other mitochondrial and cytosolic enzymes, although for the bovine enzyme the sequence was not yet extended beyond residue 72. Cys-302, a disulfiram-reactive residue in human aldehyde dehydrogenase [51], has been implicated in NAD binding [52] and is also found in all aldehyde dehydrogenases, as is Glu-268, which has been postulated to be catalytically essential [53]. Finally, Glu-487, a substituted residue in the inactive isozyme isolated from many oriental individuals [33], is conserved in all species.…”

Section: Discussionmentioning

confidence: 99%

Primary structures of rat and bovine liver mitochondrial aldehyde dehydrogenases deduced from cDNA sequences

Farrés

Guan

Weiner

1989

European Journal of Biochemistry

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The cDNA coding for rat liver mitochondrial aldehyde dehydrogenase was cloned and sequenced. It contained an open reading frame of 1557 bp. Of the deduced 519 amino acid residues, 19 were proposed to correspond to the signal peptide necessary to allow the protein to enter the mitochondria [Farrts, J., Weiner, H. (1987) Biochem. Biophys. Res. Commun. 1.50, 1083-10871. The sequence of the 500 amino acid residues comprising the mature subunit was 96% identical to that of the corresponding human liver mitochondrial enzyme. The longest cDNA isolated coding for the bovine liver enzyme contained sequence information corresponding to residues 72 -500 of the rat or human enzyme. The deduced protein sequence of these residues was 94% identical to those of the human enzyme.Over 60% of the bases in the coding region of the rat cDNA were G and C. These residues were clustered non-randomly and two potentially stable stem-loop structures could be calculated as existing in the mRNA. One would be found in the region coding for amino acid residues 64 -79; a similar secondary structure could be responsible for the existing truncated cDNAs from the bovine-cDNA libraries. Comparisons with the amino acid sequences of other aldehyde dehydrogenases support the suggestion that the NAD-binding domain may be located in the middle portion of the mature enzyme.Mammalian mitochondrial aldehyde dehydrogenase, a ubiquitous and constitutive NAD-dependent enzyme, is involved in the oxidation of biogenic aldehydes, as well as acetaldehyde formed during the metabolism of ethanol. The enzyme from most species is isolated as a tetramer of molecular mass 220 -250 kDa, resulting in a subunit molecular mass of 55 -60 kDa [l], which is larger than that of most dehydrogenases.The amino acid sequences of the human [2] and horse [3] liver mitochondrial enzymes have been published. They share 94% identity. Cytosolic enzymes from human [4] and horse [5] liver have also been sequenced and they share 91 O h identity. In contrast, mitochondrial and cytoplasmic aldehyde dehydrogenases show just 66 -68% identity.

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Active site of human liver aldehyde dehydrogenase

Cited by 96 publications

References 22 publications

Xenopus Cytosolic Thyroid Hormone-binding Protein (xCTBP) Is Aldehyde Dehydrogenase Catalyzing the Formation of Retinoic Acid

Xenopus Cytosolic Thyroid Hormone-binding Protein (xCTBP) Is Aldehyde Dehydrogenase Catalyzing the Formation of Retinoic Acid

Molecular cloning of the mitochondrial aldehyde dehydrogenase gene of Saccharomyces cerevisiae by genetic complementation

Primary structures of rat and bovine liver mitochondrial aldehyde dehydrogenases deduced from cDNA sequences

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