Skip to main content
SpringerLink
Log in

Dual function of the alcohol dehydrogenase of Drosophila melanogaster: ethanol and acetaldehyde oxidation by two allozymes ADH-71k and ADH-F

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Until recently the alcohol dehydrogenase of Drosophila melanogaster was thought to act only in the first step of primary alcohol oxidation, producing an aldehyde. Instead, acetic acid is the main product of a two-step process.

A rapid procedure was developed for the isolation and purification of two allozymes. The thermostability of the purified enzymes was found to be very different, t 1/2 at 35°C, being 45 min and 130 min for ADH-F and ADH-71k respectively.

The kinetic parameters of ethanol oxidation by the two purified allozymes were determined within physiological substrate and coenzyme ranges. The use of artificial electron acceptors has a notable influence on the ethanol oxidation: the apparent Michaelis constants increase: the oxidation rate with ADH-71k increases, whereas it decreases with ADH-F.

Purified ADH is shown to be able to catalyze the oxidation of acetaldehyde solely in the presence of NAD+, and PMS and MTT as artificial electron acceptors.

From the kinetic data the relative in vivo oxidation rates of ethanol by both ADH allozymes were calculated. ADH-F turned out to be somewhat less effective (30%–40%) than ADH-71k. The physiological consequences of these differences are discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Anderson SM, McDonald JF (1981) Effects of environmental alcohol on in vivo properties of Drosophila alcohol dehydrogenase. Biochem Genet 19:421–430

    Google Scholar 

  • Benyajati C, Place AR, Powers DA, Sofer W (1981) Alcohol dehydrogenase gene of Drosophila melanogaster: relationship of intervening sequences to functional domains in the protein. Proc Natl Acad Sci USA 78:2717–2721

    Google Scholar 

  • Berger D, Berger M, Von Wartburg JP (1974) Structural studies of human liver alcohol dehydrogenase isoenzymes. Eur J Biochem 50:215–225

    Google Scholar 

  • Chambers GK (1984) The purification and biochemical properties of alcohol dehydrogenase-“Fast (Chateau Douglas)” from Drosophila melanogaster. Biochem Genet 22:529–549

    Google Scholar 

  • Chambers GK, Laver WG, Campbell S, Gibson JB (1981) Structural analysis of an electrophoretically cryptic alcohol dehydrogenase variant from an Australian population of Drosophila melanogaster. Proc Natl Acad Sci USA 78:3103–3107

    Google Scholar 

  • Clarke B, Allendorf FW (1979) Frequency dependent selection due to kinetic differences between allozymes. Nature 279:732–733

    Google Scholar 

  • Cleland WW (1970) Steady state kinetics. In: Boyer PD (ed) The Enzymes, vol II (stud ed), Academic Press, New York, pp 1–65

    Google Scholar 

  • Crabb DW, Bosron WF, Li TK (1983) Steady state kinetic properties of purified rat liver alcohol dehydrogenase: application to predicting alcohol elimination rates in vivo. Arch Biochem Biophys 224:299–309

    Google Scholar 

  • Crow KE, Braggins TJ, Batt RD, Hardman MJ (1982) Rat liver cytosolic malate dehydrogenase: purification, kinetic properties, role in control of free cytosolic NADH concentration. J Biol Chem 257:14217–14225

    Google Scholar 

  • Daniel RM, Cowan DA, Morgan HW, Curran MP (1982) A correlation between protein thermostability and resistance to preteolysis. Biochem J 207:641–644

    Google Scholar 

  • David JR, Bocque C, Van Herrewege J, Fouillet P, Arens M-F (1978) Alcohol metabolism in Drosophila melanogaster: uselessness of the most active aldehyde oxidase produced by the Aldox locus. Biochem Genet 16:203–211

    Google Scholar 

  • De Jong F, Thörig GEW (1980) A simple test for intra-electromorphic variants of alcohol dehydrogenase in D. melanogaster. Drosophila Information Service 55:151–152

    Google Scholar 

  • Eisses KTh, Schoonen WGEJ, Scharloo W, Thörig GEW (1985) Differences in ethanol oxidation by alcohol dehydrogenase of Drosophila simulans and Drosophila melanogaster (submitted to Evolution)

  • Fersht A (1977) Enzyme structure and mechanism. WHF Freeman & Co, Reading and San Francisco

    Google Scholar 

  • Geer BW, McKechnie SW, Langevin ML (1983) Regulation of sn-glycerol-3-phosphate dehydrogenase in Drosophila melanogaster larvae by dietary ethanol and sucrose. J Nutrition 113:1632–1642

    Google Scholar 

  • Gibson JB, Oakeshott JG (1982) Tests of the adaptive significance of the alcohol dehydrogenase polymorphism in Drosophila melanogaster: paths, pitfalls and prospects. In: Barker JSF, Starmer WT (eds) Ecological genetics and evolution. The cactus-yeast-Drosophila model system. Academic Press Australia, North Ryde, NSW, pp 291–306

    Google Scholar 

  • Hall JG, Koehn RK (1983) The evolution of enzyme catalytic efficiency and adaptive inference from steady state kinetic data. Evol Biol 16:53–96

    Google Scholar 

  • Heinstra PWH, Eisses KTh, Schoonen WGEJ, Aben W, De Winter AJ, Van der Horst DJ, Van Marrewijk WJA, Beenakkers AMTh, Scharloo W, Thörig GEW (1983) A dual function of alcohol dehydrogenase in Drosophila. Genetica 60:129–137

    Google Scholar 

  • Heinstra PWH, Eisses KTh, Scharloo W, Thörig GEW (1985) Metabolism of secondary alcohols in Drosophila melanogaster: Effects on alcohol dehydrogenase. (Submitted to Biochem Genet)

  • Hovik R, Winberg J-O, McKinley-McKee JS (1984) Drosophila melanogaster alcohol dehydrogenase: substrate stereospecificity of the Adh-F alleloenzyme. Insect Biochem 14:345–351

    Google Scholar 

  • McElfresh KC, McDonald JF (1983) The effect of alcohol stress on Nicotinamide Adenine Dinucleotide (NAD+) levels in Drosophila. Biochem Genet 21:365–374

    Google Scholar 

  • McKechnie SW, Morgan P (1982) Alcohol dehydrogenase polymorphism of Drosophila melanogaster: aspects of alcohol and temperature variation in the larval environment. Aust J Biol Sci 35:85–93

    Google Scholar 

  • McKechnie SW, Geer BW (1984) Regulation of alcohol dehydrogenase in Drosophila melanogaster by dietary alcohol and carbohydrate. Insect Biochem 14:231–242

    Google Scholar 

  • Middleton RJ, Kacser H (1983) Enzyme variation, metabolic flux and fitness: alcohol dehydrogenase in Drosophila melanogaster. Genetics 105:633–650

    Google Scholar 

  • Papel I, Henderson M, Van Herrewege J, David J, Sofer W (1979) Drosophila alcohol dehydrogenase activity in vivo and in vitro. Effects of acetone feeding. Biochem Genet 17:553–563

    Google Scholar 

  • Perutz MF (1980) What the abnormal hemoglobins teach us about the folding problem. In: Jaenicke R (ed) Protein Folding. Elsevier, North Holland Biomed Press, Amsterdam, pp 317–322

    Google Scholar 

  • Retzios A, Thatcher DR (1979) Chemical basis of the electrophoretic variation observed at the alcohol dehydrogenase locus of Drosophila melanogaster. Biochimie 61:701–704

    Google Scholar 

  • Rudolph FB, Fromm HJ (1979) Plotting methods for analyzing enzyme rate data. In: Purich DL (ed) Methods in enzymology, vol 63A, Academic Press, New York, pp 138–159

    Google Scholar 

  • Scharloo W, Van Dijken FR, Hoorn AJW, DeJong G, Thörig GEW (1977) Functional aspects of genetic variation. In: Christiansen FB, Fenchel TM (eds) Measuring selection in natural populations. Springer, Berlin Heidelberg New York, pp 135–145

    Google Scholar 

  • Schwarz M, Sofer W (1976) Diet-induced alterations in distribution of multiple forms of alcohol dehydrogenase in Drosophila. Nature 263:129–131

    Google Scholar 

  • Thörig GEW, Schoone AA, Scharloo W (1975) Variation between electrophoretically identical alleles at the alcohol dehydrogenase locus in Drosophila melanogaster. Biochem Genet 13: 721–731

    Google Scholar 

  • van Delden W (1982) The alcohol dehydrogenase polymorphism in Drosophila melanogaster. Selection at an enzyme locus. Evol Biol 15:187–222

    Google Scholar 

  • Vilageliu Arqués LI, González Duarte R (1980) Effect of ethanol and isopropanol on the activity of ADH, viability and life span in Drosophilas melanogaster and Drosophila funebris. Experientia 36:828–830

    Google Scholar 

  • Wills C, Jörnvall H (1979) Amino acid substitutions in two functional mutants of yeast alcohol dehydrogenase. Nature 279:734–736

    Google Scholar 

  • Winberg J-O, Thatcher DR, McKinley-McKee JS (1982) Alcohol dehydrogenase from the fruitfly Drosophila melanogaster: substrate specificity of the alleloenzymes ADH-S and ADH-UF. Biochem Biophys Acta 704:7–16

    Google Scholar 

  • Zera AJ, Koehn RK, Hall JG (1985) Allozymes and biochemical adaptation. In: Kerkut GK, Gilbert LI (eds) Comprehensive insect physiology, Biochemistry and Pharmacology, vol 10. Pergamon Press, New York

    Google Scholar 

Download references

Author information

Author notes

  1. Willem G. E. J. Schoonen

    Present address: Zoological Laboratory, Rijksuniversiteit Utrecht, Padualaan 8, NL-3584 CH, Utrecht, The Netherlands

Authors and Affiliations

  1. Department of Population and Evolutionary Biology, Rijksuniversiteit at Utrecht, Padualaan 8, NL-3584 CH, Utrecht, The Netherlands

    Karel Th. Eisses, Willem G. E. J. Schoonen, Wim Aben, Willem Scharloo & George E. W. Thörig

Authors
  1. Karel Th. Eisses
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Willem G. E. J. Schoonen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wim Aben
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Willem Scharloo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. George E. W. Thörig
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by M.M. Green

Rights and permissions

Reprints and permissions

About this article

Cite this article

Eisses, K.T., Schoonen, W.G.E.J., Aben, W. et al. Dual function of the alcohol dehydrogenase of Drosophila melanogaster: ethanol and acetaldehyde oxidation by two allozymes ADH-71k and ADH-F. Molec Gen Genet 199, 76–81 (1985). https://doi.org/10.1007/BF00327513

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00327513

Keywords

Search

Navigation