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

Abstract

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.

Proteome analysis of rat liver mitochondria reveals a possible compensatory response to endotoxic shock

“…3). Table 1) [20]. We have determined three different ALDH1-patterns depending on the distribution of four isoforms of this enzyme (see Fig.…”

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

“…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).…”

“…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.…”

Structural features of stomach aldehyde dehydrogenase distinguish dimeric aldehyde dehydrogenase as a ‘variable’ enzyme ‘Variable’ and ‘constant’ enzymes within the alcohol and aldehyde dehydrogenase families

“…Several peptides wills identical N.termlnal structures elute al more titan one position, presumably becattse of separate secondary modifications or different C,terntinal cleavages, utilizing arylamine membranes for C.~ermina~ coupli.lL Results obtained were correlated wttlt known ~tldehyde dehydrogenase structures [19] and with the different ~voh.donary rates for mitochondrial aldehyde dehydrogcnase, cytosolie class I and Ill alcohol dehydro. genases, a,td sorbitol dehydrogenase, reeenlly established from the structures of those rat/human enzymes (el, [13][14][15][16][17][18][19],),…”

“…Only a brief report [10] on limited data without further details exists. Furthermore, knowledge on the structural variation of dimeric aldehyde dehydrogenase is of special interest, since other alcohol and aldehyde dehydrogenases differ widely in being either fairly 'constant' (class III alcohol dehydrogenase and mitochondrial aldehyde dehydrogenase [13][14][15]) or 'variable' (class I alcohol dehydrogenase, sorbitol dehydrogenase and cytosolic aldehyde dehydrogenase [13,16]). …”