SEARCH

SEARCH BY CITATION

Abstract

  1. Top of page
  2. Abstract
  3. References

Human stomach mucosa contains a characteristic alcohol dehydrogenase (ADH) enzyme, σσ-ADH. Its cDNA has been cloned from a human stomach library and sequenced. The deduced amino acid sequence shows 59–70% identities with the other human ADH classes, demonstrating that the stomach enzyme represents a distinct structure, constituting class IV, coded by a separate gene, ADH7. The amino acid identity with the rat stomach class IV ADH is 88%, which is intermediate between constant and variable dehydrogenases. This value reflects higher conservation than for the classical liver enzymes of class I, compatible with a separate functional significance of the class IV enzyme. Its enzymic features can be correlated with its structural characteristics. The residues lining the substrate-binding cleft are bulky and hydrophobic, similar to those of the class I enzyme; this explains the similar specificity of both classes, compatible with the origin of class IV from class I. Position 47 has Arg, in contrast to Gly in the rat class IV enzyme, but this Arg is still associated with an extremely high activity (kcat= 1510 min-1) and weak coenzyme binding (KiaNAD+= 1.6 mM). Thus, the strong interaction with coenzyme imposed by Arg47 in class I is probably compensated for in class IV by changes that may negatively affect coenzyme binding: Glu230, His271, Asn260, Asn261, Asn363. The still higher activity and weaker coenzyme binding of rat class IV (kcat= 2600 min-1, KiaNAD = 4 mM) can be correlated to the exchanges to Gly47, Gln230 and Tyr363. An important change at position 294, with Val in human and Ala in rat class IV, is probably responsible for the dramatic difference in Km values for ethanol between human (37 mM) and rat (2.4 M) class IV enzymes.

Abbreviation
ADH

alcohol dehydrogenase

Enzyme
 

Alcohol dehydrogenase (EC 1.1.1.1)

References

  1. Top of page
  2. Abstract
  3. References
  • Benson, S. A. & Taylor, R. K. (1984) A rapid small scale procedure for isolation of phage lambda DNA, Biotechniques 2, 126127.
  • Berget, S. M. (1984) Are U4 small nuclear ribonucleoproteins involved in polyadenylation? Nature 309, 179181.
  • Boleda, M. D., Saubi, N., Farrés, J. & Parés, X. (1993) Physiological substrates for rat alcohol dehydrogenase classes: Aldehydes of lipid peroxidation, ω-hydroxyfatty acids and retinoids, Arch. Biochem. Biophys. 307, 8590.
  • Bosron, W. F., Li, T.-K., Dafeldecker, W. P. & Vallee, B. L. (1979) Human liver n-alcohol dehydrogenase: kinetic and molecular properties, Biochemistry 18, 11011105.
  • Bosron, W. F., Magnes, L. J. & Li, T.-K. (1983) Kinetic and electrophoretic properties of native and recombined isoenzymes of human liver alcohol dehydrogenase, Biochemistry 22, 18521857.
  • Chen, C.-S. & Yoshida, A. (1991) Enzymatic properties of the protein encoded by newly cloned human alcohol dehydrogenase ADH6 gene, Biochem. Biophys. Res. Commun. 181, 743747.
  • Danielsson, O., Atrian, S., Luque, T., Hjelmqvist, L., Gonzàlez-Duarte, R. & Jörnvall, H. (1994) Fundamental molecular differences between alcohol dehydrogenase classes. Drosophila octanol dehydrogenase is a class III alcohol dehydrogenase establishing molecular patterns and a continuous distribution of this zinc enzyme family, Proc. Natl Acad. Sci. USA 91, 49804984.
  • Devereux, J., Haeberli, P. & Smithies, O. (1984) A comprehensive set of sequence analysis programs for the VAX, Nucleic Acids Res. 12, 387395.
  • Egestad, B., Estonius, M., Danielsson, O., Persson, B., Cederlund, E., Kaiser, R., Holmquist, B., Vallee, B., Parés, X., Jeffery, J. & Jörnvall, H. (1990) Fast atom bombardment mass spectrometry and chemical analysis in determinations of acyl-blocked protein structures, FEBS Lett. 269, 194196.
  • Eklund, H., Samama, J. P. & Jones, T. A. (1984) Crystallographic investigations of nicotinamide adenine dinucleotide binding to horse liver alcohol dehydrogenase, Biochemistry 23, 59825996.
  • Eklund, H., Müller-Wille, P., Horjales, E., Futer, O., Holmquist, B., Vallee, B. L., Höög, J.-O., Kaiser, R. & Jörnvall, H. (1990) Comparison of three classes of human liver alcohol dehydrogenase. Emphasis on different substrate-binding pockets, Eur. J. Biochem. 193, 303310.
  • Estonius, M., Danielsson, O., Karlsson, C., Persson, H., Jörnvall, H. & Höög, J.-O. (1993) Distribution of alcohol and sorbitol dehydrogenases. Assessment of mRNA species in mammalian tissues, Eur. J. Biochem. 215, 497503.
  • Flinta, C., Persson, B., Jörnvall, H. & von Heijne, G. (1986) Sequence determinants of cytosolic N-terminal protein processing, Eur. J. Biochem. 154, 193196.
  • Frezza, M., di Padova, C., Pozzato, G., Terpin, M., Baraona, E. & Lieber, C. S. (1990) High blood alcohol levels in women. The role of decreased gastric alcohol dehydrogenase activity and first-pass metabolism, N. Engl. J. Med. 322, 9599.
  • Friedman, K. D., Rosen, N. L., Newman, P. J. & Montgomery, R. R. (1988) Enzymatic amplification of specific cDNA inserts from lambda gt11 libraries, Nucleic Acids Res. 16, 8718.
  • Hedén, L.-O., Höög, J.-O., Larsson, K., Lake, M., Lagerholm, E., Holmgren, A., Vallee, B. L., Jörnvall, H. & von Bahr-Lindström, H. (1986) cDNA clones coding for the β-subunit of human liver alcohol dehydrogenase have differently sized 3′-non-coding regions, FEBS Lett. 194, 327332.
  • Holmes, R. S. (1988) Alcohol dehydrogenases and aldehyde dehydrogenases of anterior eye tissues from humans and other mammals, in Biomedical and social aspects of alcohol and alcoholism (Kuriyama, K., Takada, A. & Ishii, H., eds) pp. 5157, Elsevier Science Publishers, New York .
  • Höög, J.-O., Hedén, L.-O., Larsson, K., Jörnvall, H. & von Bahr Lindström, H. (1986) The γ1 and γ2 subunits of human liver alcohol dehydrogenase. cDNA structures, two amino acid replacements and compatibility with changes in the enzymatic properties, Eur. J. Biochem. 159, 215218.
  • Höög, J.-O., von Bahr-Lindström, H., Hedén, L.-O., Holmquist, B., Larsson, K., Hempel, J., Vallee, B. L. & Jörnvall, H. (1987) Structure of the class II enzyme of human liver alcohol dehydrogenase: Combined cDNA and protein sequence determination of the π subunit, Biochemistry 26, 19261932.
  • Hurley, T. D., Edenberg, H. J. & Bosron, W. F. (1990) Expression and kinetic characterization of variants of human β1β1 alcohol dehydrogenase containing substitutions at amino acid 47, J. Biol. Chem. 265, 1636616372.
  • Hurley, T. D., Bosron, W. F., Hamilton, J. A. & Amzel, L. M. (1991) Structure of human β1β1 alcohol dehydrogenase: Catalytic effects of non-active-site substitutions, Proc. Natl Acad. Sci. USA 88, 81498153.
  • Jörnvall, H., Persson, B. & Jörnvall, H. (1993) Variability patterns of dehydrogenases versus peptide hormones and proteases/anti-proteases, FEBS Lett. 335, 6972.
  • Jörnvall, H. & Höög, J.-O. (1994) Nomenclature of alcohol dehydrogenases, Alcohol Alcohol., in the press.
  • Julià, P., Farrés, J. & Parés, X. (1987) Characterization of three isoenzymes of rat alcohol dehydrogenase. Tissue distribution and physical and enzymatic properties, Eur. J. Biochem. 162, 179189.
  • Koivusalo, M., Baumann, M. & Uotila, L. (1989) Evidence for the identity of glutathione-dependent formaldehyde dehydrogenase and class III alcohol dehydrogenase, FEBS Lett. 257, 105109.
  • Kozak, M. (1987) An analysis of 5′-non coding sequences from 699 vertebrate messenger RNAs, Nucleic Acids Res. 15, 81258148.
  • Maniatis, T., Fritsch, E. F. & Sambrook, J. (1982) Molecular cloning: a laboratory manual, Cold Spring Harbor Laboratory, Cold Spring Harbor NY .
  • McLauchlan, J., Gaffney, D., Whitton, J. L. & Clements, J.-B. (1985) The consensus sequence YGTGTTYY located downstream from the AATAAA signal is required for efficient formation of mRNA 3′ termini, Nucleic Acids Res. 13, 13471368.
  • Moreno, A. & Parés, X. (1991) Purification and characterization of a new alcohol dehydrogenase from human stomach, J. Biol. Chem. 266, 11281133.
  • Moreno, A., Parés, A., Ortiz, J., Enríquez, J. & Parés, X. (1994) Alcohol dehydrogenase from human stomach. Variability in normal mucosa and effect of age, sex, ADH3 phenotype and gastric region, Alcohol Alcohol., in the press.
  • Parés, X., Moreno, A., Cederlund, E., Höög, J.-O. & Jörnvall, H. (1990) Class IV mammalian alcohol dehydrogenase. Structural data of the rat stomach enzyme reveal a new class well separated from those already characterized, FEBS Lett. 277, 115118.
  • Parés, X., Cederlund, E., Moreno, A., Saubi, N., Höög, J.-O. & Jörnvall, H. (1992) Class IV alcohol dehydrogenase (the gastric enzyme). Structural analysis of human σσ-ADH reveals class IV to be variable and confirms the presence of a fifth mammalian alcohol dehydrogenase class, FEBS Lett. 303, 6972.
  • Parés, X., Cederlund, E., Moreno, A., Hjelmqvist, L., Farrés, J. & Jörnvall, H. (1994) Mammalian class IV alcohol dehydrogenase (stomach alcohol dehydrogenase): Structure, origin, and correlation with enzymology, Proc. Natl Acad. Sci. USA 91, 18931897.
  • Pearson, W. R. & Lipman, D. J. (1988) Improved tools for biological sequence analysis, Proc. Natl Acad. Sci. USA 85, 24442448.
  • Persson, B., Bergman, T., Keung, W. M., Waldenström, U., Holmquist, B., Vallee, B. L. & Jörnvall, H. (1993) Basic features of class-I alcohol dehydrogenase: variable and constant segments coordinated by inter-class and intra-class variability. Conclusions from characterization of the alligator enzyme, Eur. J. Biochem. 216, 4956.
  • Proudfoot, N. J. & Brownlee, G. G. (1976) 3′ Non-coding region sequences in eukaryotic messenger RNA, Nature 263, 211214.
  • Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989) Molecular cloning: a laboratory manual, 2nd edn, Cold Spring Harbor Laboratory, Cold Spring Harbor NY .
  • Sanger, F., Nicklen, S. & Coulson, A. R. (1977) DNA sequencing with chain-terminating inhibitors, Proc. Natl Acad. Sci. USA 74, 54635467.
  • Segel, I. H. (1975) Enzyme kinetics, John Wiley & Sons, New York .
  • Sharma, C. P., Fox, E. A., Holmquist, B., Jörnvall, H. & Vallee, B. L. (1989) cDNA sequence of human class III alcohol dehydrogenase, Biochem. Biophys. Res. Commun. 164, 631637.
  • Stone, C. L., Thomasson, H. R., Bosron, W. F. & Li, T.-K. (1993) Purification and partial amino acid sequence of a high-activity human stomach alcohol dehydrogenase, Alcohol. Clin. Exp. Res. 17, 911918.
  • Vallee, B. L. & Bazzone, T. J. (1983) Isozymes of human liver alcohol dehydrogenase, Isozymes Curr. Top. Biol. Med. Res. 8, 219244.
  • Yasunami, M., Chen, C.-S. & Yoshida, A. (1991) A human alcohol dehydrogenase gene (ADH 6) encoding an additional class of isozyme, Proc. Natl Acad. Sci. USA 88, 76107614.
  • Yin, S. J., Bosron, W. E., Magnes, L. J. & Li, T.-K. (1984) Human liver alcohol dehydrogenase: purification and kinetic properties of the β2β2, β1β2, αβ2, and β2γ1‘Oriental’ isoenzymes, Biochemistry 23, 58475853.
  • Yin, S.-J., Wang, M.-F., Liao, C.-S., Chen, C.-M. & Wu, C.-W. (1990) Identification of a human stomach alcohol dehydrogenase with distinctive kinetic properties, Biochem. Int. 22, 829835.
  • Yin, S.-J., Vagelopoulos, N., Wang, S.-L. & Jörnvall, H. (1991) 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, FEBS Lett. 283, 8588.
  • Zheng, Y.-W., Bey, M., Liu, H. & Felder, M. R. (1993) Molecular basis of the alcohol dehydrogenase-negative deer mouse. Evidence for deletion of the gene for class I enzyme and identification of a possible new enzyme class, J. Biol. Chem. 268, 2493324939.