Journal of Comparative Physiology B

, Volume 161, Issue 2, pp 207–212 | Cite as

Differential survival of Venus gallina and Scapharca inaequivalvis during anoxic stress: Covalent modification of phosphofructokinase and glycogen phosphorylase during anoxia

  • S. P. J. Brooks
  • A. de Zwaan
  • G van den Thillart
  • O. Cattani
  • P. Cortesi
  • K. B. Storey


Biochemical mechanisms underlying anaerobiosis were assessed in two Mediterranean bivalve species, Scapharca inaequivalvis and Venus gallina, with widely differing tolerances for oxygen lack. These species displayed LT50 values for anoxic survival at 17–18°C of 17 and 4 d, respectively. Succinate and alanine were the major products of 24 h anaerobic metabolism in both species but only S. inaequivalvis further metabolized succinate to propionate. Both species reduced metabolic rate while anoxic but metabolic arrest was more pronounced in S. inaequivalvis. Calculated ATP turnover rate (MATP) during exposure to N2-bubbled seawater was only 4.51% of the aerobic rate in S. inaequivalvis but was 12.68% in V. gallina. To counteract a greater load of acid end products, V. gallina foot showed a significantly greater buffering capacity, 23.38±0.20 slykes, compared to 19.6±0.79 slykes in S. inaequivalvis. The two species also differed distinctly in the enzymatic regulation of anaerobiosis. In V. gallina anoxia exposure caused only a small change in PFK kinetic parameters (a decrease in Ka AMP) and had no effect on glycogen phosphorylase. By contrast, S. inaequivalvis foot showed a strong modification of enzyme properties in anoxia. The percentage of glycogen phosphorylase in the a form dropped significantly only in S. inaequivalvis. Other changes included alterations in the properties of PFK leading to a less active enzyme form in anoxia. Compared to the aerobic enzyme form, PFK from anoxic foot showed a reduced affinity for fructose-6-P (Km increased 2.4-fold), greater inhibition by ATP (I50 decreased 6.8-fold), and an increase in sensitivity to AMP activation (Ka decreased by 50%). These enzyme changes appear to be key to a glycolytic rate depression during anaerobiosis in S. inaequivalvis foot muscle.

Key words

Anaerobiosis Glycolytic control Phosphofructokinase Glycogen phosphorylase Molluscs 



ethylenediaminetetraacetic acid


ethyleneglycol-bis-(2-aminoethyl)-tetraacetic acid






Activation constant (concentration of AMP required to increase the reaction to twice the rate it shows in the absence of AMP)


ATP turnover rate


inorganic phosphate


Perchloric acid




Trichloroacetic acid


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  1. Castellini MA, Somero GN (1981) Buffering capacity of vertebrate muscle: correlations with potentials for anaerobic function. J Comp Physiol B 143:191–198Google Scholar
  2. Eberlee JC, Storey KB (1984) Buffering capacities of the tissues of marine molluscs. Physiol Zool 57:567–572Google Scholar
  3. Ghisotti F, Rinaldi E (1976) Osservazioni sulla popolazione di Scapharca insediatasi in questi ultimi anni su un tratto di littorale romagnolo. Conchiglie 12:183–185Google Scholar
  4. Helmerhorst E, Stokes GB (1983) Microcentrifuge desalting: A rapid, quantitative method for desalting small amounts of protein. Anal Biochem 104:130–135Google Scholar
  5. Holwerda DA, Veenhof PR, van Heugten HAA, Zandee DI (1983) Modification of mussel pyruvate kinase during anaerobiosis and after temperature acclimation. Mol Physiol 3:225–234Google Scholar
  6. Hochachka PW (1980) Living without oxygen. Harvard University press, Cambridge MAGoogle Scholar
  7. Isani G (1987) Anaerobic metabolism in molluscan bivalves of the Adriatic Sea. PhD. Thesis, Universitá degli Studi di Bologna, Bologna ItalyGoogle Scholar
  8. Isani G, Cattani O, Carpené E, Tacconi S, Cortesi P (1989) Energy metabolism during anaerobiosis and recovery in the posterior adductor muscle of the bivalve Scapharca inaequivalvis (Bruguiere). Comp Biochem Physiol B 93:193–200Google Scholar
  9. Kluytmans JH, de Bont AMT, Kruitwagen ECJ, Ravestein HJL, Veenhof PR (1983) Anaerobic capacities and anaerobic energy production of some Mediterranean bivalves. Comp Biochem Physiol B 75:171–179Google Scholar
  10. Korycan SA (1984) A study of the metabolic adaptations of a bivalve mollusc to environmental stresses. M.Sc. Thesis, Carleton University, Ottawa, CanadaGoogle Scholar
  11. Kreutzer U, Siegmund B, Grieshaber MK (1985) Role of coupled substrates and alternative end products during hypoxia in marine invertebrates. Mol Physiol 8:371–392Google Scholar
  12. Livingstone DL, Zwaan A de (1983) Carbohydrate metabolism in gastropods. In: Wilbur KM (ed) The Mollusca, vol. 1. Academic Press, New York, pp 177–242Google Scholar
  13. Michaelidis B, Gaitanaki C, Beis I (1988) Modification of pyruvate kinase from the foot muscle of Patella caerulea (L.) during anaerobiosis. J Exp Zool 248:264–271Google Scholar
  14. Morris GM, Baldwin J (1984) pH buffering capacity of invertebrate muscle: correlations with anaerobic muscle work. Mol Physiol 5:61–70Google Scholar
  15. Plaxton WC, Storey KB (1984) Phosphorylation in vivo of redmuscle pyruvate kinase from the channelled whelk, Busycotypus canaliculatum, in response to anoxic stress. Eur J Biochem 143:267–272Google Scholar
  16. Plaxton WC, Storey B (1985) Tissue specific isozymes of pyruvate kinase in the channelled whelk Busycotypus canaliculatum: enzyme modification in response to environmental anoxia. J Comp Physiol B 155:291–296Google Scholar
  17. Regione Emilia Romagna, Assessorato ambiente e difesa del suolo (1985) Eutrofizzazione delle acque costiere dell'Emilia-Romagna, Italia. Nespoli G (coordinator).Google Scholar
  18. Regione Emilia Romagna Assessorato ambiente e difesa del suolo (1986) Eutrofizzazione delle acque costiere dell'Emilia-Romagna, Italia. Nespoli G (coordinator).Google Scholar
  19. Schaftingen E van (1984) d-Fructose-2,6-bisphosphate. In: Bergmeyer HU (ed) Methods of enzymatic analysis, 3rd edition. Verlag Chemie, Weinheim, pp 335–341Google Scholar
  20. Slyke DD van (1922) On the measurement of buffer values and on the relationship of buffer value to the dissociation constant of the buffer and the concentration and reaction of the buffer solution. J Biol Chem 52:525–570Google Scholar
  21. Storey KB (1984) Phosphofructokinase from foot muscle of the whelk, Busycotypus canaliculatum: evidence for covalent modification of the enzyme during anaerobiosis. Arch Biochem Biophys 235:665–672Google Scholar
  22. Storey KB (1985) A re-evaluation of the Pasteur effect: new mechanisms in anaerobic metabolism. Mol Physiol 8:439–461Google Scholar
  23. Storey KB (1988a) Suspended animation: the molecular basis of metabolic rate depression. Can J Zool 66:124–132Google Scholar
  24. Storey KB (1988b) Mechanisms of glycolytic control during facultative anaerobiosis in a marine mollusc: tissue specific analysis of glycogen phosphorylase and fructose-2,6-bisphosphate. Can J Zool 66:1767–1771Google Scholar
  25. Whitwam RE, Storey KB (1990) Organ specific regulation of phosphofructokinase during facultative anaerobiosis in the marine whelk, Busycotypus canaliculatum. Can J Zool (in press)Google Scholar
  26. Zandee DI, de Zwaan A, Cortesi P, Cattani O (1988) Biochemical parameters to judge effect of environmental variables in energy metabolism. Proceedings of the International Union of Biological Science (2nd Int Congress). Abstract No. 618. Baton Rouge, Louisiana, USAGoogle Scholar
  27. Zurburg W, Kluytmans JH (1980) Organ specific changes in energy metabolism due to anaerobiosis in the sea mussel Mytilus edulis L. Comp Biochem Physiol 67B:317–322Google Scholar
  28. Zwaan A de (1983) Carbohydrate metabolism in bivalves. In: Wilbur KM (ed) The Mollusca, vol. 1. Academic Press, New York, pp 137–175Google Scholar
  29. Zwaan A de, Bont AMT de, Verhoeven A (1982) Anaerobic energy metabolism in isolated adductor muscle of the sea mussel Mytilus edulis L. J Comp Physiol 149:137–143Google Scholar
  30. Zwaan A de, Cortesi P, Thillart G van den, Brooks S, Storey KB, Roos J, Lieshout G van, Cattani O, Vitali G (1990) Energy metabolism of bivalves at reduced oxygen tensions. Proceedings Marine Coastal Eutrophication. Int. Conf. Bologna (I) (in press)Google Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • S. P. J. Brooks
    • 1
  • A. de Zwaan
    • 1
  • G van den Thillart
    • 1
  • O. Cattani
    • 1
  • P. Cortesi
    • 1
  • K. B. Storey
    • 1
  1. 1.Consorzio di Studi, Ricerche ed Interventi sulle Risorse MarineCesenatico, FOItaly

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