Mammalian alcohol dehydrogenases of separate classes: intermediates between different enzymes and intraclass isozymes.

Jörnvall, Hans; Höög, Jan‐Olov; Bahr‐Lindström, Hedvig von; Vallée, Bert L.

doi:10.1073/pnas.84.9.2580

Cited by 72 publications

(34 citation statements)

References 30 publications

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“…3). These structural results support the classification of the class 111 enzymes as a different type of alcohol dehydrogenase derived from an early duplication [8], in agreement cLass1 133 differences ~(~~~ 164% identity) and y z subunits are also known [7] but not included in the figure since they only affect values marginally) Fig. 3), suggesting more strict structure/function relationships for the class I11 enzyme type.…”

Section: Overall Propertiessupporting

confidence: 66%

“…strands in the coenzyme-binding domain (PA-BF) [4, 51 present few and highly conservative exchanges in the same manner as found in the class 1/11 comparison [8]. A similar distribution of conservation between the human class I isozymes has also been reported [7].…”

Section: Coenzyme-binding Region and The Segment Involved In Subunitmentioning

confidence: 56%

“…Class I ezymes are very sensitive to pyrazole inhibition and have a considerable affinity for ethanol (human ADH isozymes with a, 8, y subunits and rat ADH-3) ; class I11 alcohol dehydrogenase is virtually insensitive to pyrazole and has a very low affinity for ethanol (human x-ADH [18] and rat ADH-2 [17]); while the class I1 enzymes have intermediate properties (human 71-ADH and rat ADH-1). Structural studies are required to understand these and other properties.…”

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confidence: 99%

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Rat liver alcohol dehydrogenase of class III

Julià

Parés

Jörnvall

1988

European Journal of Biochemistry

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The amino acid sequence of alcohol dehydrogenase of class I11 from rat liver (the enzyme ADH-2) has been determined. This type of structure is quite different from those of both the class I and the class I1 alcohol dehydrogenases. The rat class I11 structure differs from the rat and human class I structures by 133 -138 residues (exact value depending on species and isozyme type); and from that of human class I1 by 132 residues. In contrast, the rat/human species difference within the class I11 enzymes is only 21 residues. The protein was carboxymethylated with iod0[2~~C]acetate, and cleaved with CNBr and proteolytic enzymes. Peptides purified by exclusion chromatography and reverse-phase high-performance liquid chromatography were analyzed by degradation with a gas-phase sequencer and with the manual 4-N,N-dimethylaminoazobenzene-4'-isothiocyanate double-coupling method.The protein chain has 373 residues with a blocked N terminus. No evidence was obtained for heterogeneity. The rat ADH-2 enzyme of class I11 contains an insertion of Cys at position 60 in relation to the class I enzymes, while the latter alcohol dehydrogenase in rat (ADH-3) has another Cys insertion (at position 111) relative to ADH-2. The structure deduced explains the characteristic differences of the class I11 alcohol dehydrogenase in relation to the other classes of alcohol dehydrogenase, including a high absorbance, an anodic electrophoretic mobility and special kinetic properties. The main amino acid substitutions are found in the catalytic domain and in the subunit interacting segments of the coenzyme-binding domain, the latter explaining the lack of hybrid dimers between subunits of different classes. Several substitutions provide an enlarged and more hydrophilic substrate-binding pocket, which appears compatible with a higher water content in the pocket and hence could possibly explain the higher K , for all substrates as compared with the corresponding values for the class I enzymes. Finally the class I11 structure supports evolutionary relationships suggesting that the three classes constitute clearly separate enzymes within the group of mammalian zinc-containing alcohol dehydrogenases.Mammalian alcohol dehydrogenases (ADH) are zinc-containing enzymes with known relationships to similar enzymes in yeast, plants and other organisms [l -31. The horse enzyme constitutes the crystallographically investigated model enzyme [4] (recent review in [5]). The human enzymes form a complex system of three classes [6], which have all recently been characterized (summaries in [7 -91).

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Section: Overall Propertiessupporting

confidence: 66%

Section: Coenzyme-binding Region and The Segment Involved In Subunitmentioning

confidence: 56%

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confidence: 99%

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Rat liver alcohol dehydrogenase of class III

Julià

Parés

Jörnvall

1988

European Journal of Biochemistry

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“…Three classes of human alcohol dehydrogenase, I-III, were distinguished early (1) and have been characterized structurally as separate forms related by gene duplications that are intermediate between those giving rise to isozymes and those giving rise to entirely different enzymes (2). Additional classes, IV and V (3)(4)(5)(6)(7)(8), of these medium-chain alcohol dehydrogenases and further isozymes in some species (9,10) have since been added.…”

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confidence: 99%

Mammalian class IV alcohol dehydrogenase (stomachalcohol dehydrogenase): structure, origin, and correlation withenzymology.

Parés

Cederlund

Moreno

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

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The structure of a mammalian class IV alcohol dehydrogenase has been determined by peptide analysis of the protein isolated from rat stomach. The structure indicates that the enzyme constitutes a separate alcohol dehydrogenase class, in agreement with the distinct enzymatic properties; the class IV enzyme is somewhat closer to class I (the "classical" liver alcohol dehydrogenase; -68% residue identities) than to the other classes (II, m, and V; -60% residue identities), suggesting that class IV might have originated through duplication of an early vertebrate class I gene. The activity of the class IV protein toward ethanol is even higher than that of the classical liver enzyme. Both Km and kalt values are high, the latter being the highest of any class characterized so far. Structurally, these properties are correlated with replacements at the active site, affecting both substrate and coenzyme binding. In particular, Ala-294 (instead of valine) results in increased space in the middle section of the substrate cleft, Gly-47 (instead of a basic residue) results in decreased charge interactions with the coenzyme pyrophosphate, and Tyr-363 (instead of a basic residue) may also affect coenzyme binding. In combination, these exchanges are compatible with a promotion of the off dissociation and an increased turnover rate. In contrast, residues at the inner part of the substrate cleft are bulky, accounting for low activity toward secondary alcohols and cyclohexanol. Exchanges at positions 259-261 involve minor shifts in glycine residues at a reverse turn in the coenzyme-binding fold. Clearly, class IV is distinct in structure, ethanol turnover, stomach expression, and possible emergence from class I.

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“…This segment has been noted in comparisons of classes [18] and found to account for the failure of dimeric hybrids to form with subunits of different classes. As evidenced by the present data, this segment is also variable within a class.…”

Section: Segment Architecture Of Class 1 Alcohol Dehydrogenasementioning

confidence: 99%

Basic features of class‐I alcohol dehydrogenase: variable and constant segments coordinated by inter‐class and intra‐class variability

Persson

Bergman

Keung

et al. 1993

European Journal of Biochemistry

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The enzymatic and structural properties of alligator liver alcohol dehydrogenase have been determined. Aliphatic and alicyclic alcohols serve as substrates for this first reptilian form of the enzyme characterized, with K,, values decreasing rapidly from methanol to hexanol, as for the human class I enzymes, and a K,, of 1.2 mM for ethanol at pH 9.9. The N-terminus of the 374-residue protein chain is acetyl-blocked.The enzyme is related in descending order to class I > 111 > V > I1 of the structurally characterized mammalian alcohol dehydrogenases. This observation is compatible with the presence of a I/III ancestral line. Differences of the enzyme classes exceed those of the species, suggesting an early origin of the classes. Within its enzyme class, the reptilian protein is most closely related to the avian form (82% residue identities), and is closer to the human than to the amphibian form (76%, versus 69%, respectively). This establishes class I alcohol dehydrogenase as an enzyme having fairly constant rate of change during much of vertebrate evolution, -10% residue differences/lOO million years of separation between pairs compared.Residues interacting with the substrate and coenzyme are largely conserved. In the alligator enzyme, there are only four replacements in the substrate pocket compared with the human class I y subunit, and those are not known to have functional roles. These properties account for the kinetic parameters, and suggest distinct metabolic functions for the class I enzyme in vertebrates.Comparisons of the enzymes of the different vertebrate lines reveal that segment patterns are characteristic features of the class I enzymes.

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Mammalian alcohol dehydrogenases of separate classes: intermediates between different enzymes and intraclass isozymes.

Cited by 72 publications

References 30 publications

Rat liver alcohol dehydrogenase of class III

Rat liver alcohol dehydrogenase of class III

Mammalian class IV alcohol dehydrogenase (stomachalcohol dehydrogenase): structure, origin, and correlation withenzymology.

Basic features of class‐I alcohol dehydrogenase: variable and constant segments coordinated by inter‐class and intra‐class variability

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