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

SummaryBiochemical 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.

[1]  J. Baldwin,et al.  pH buffering capacity of invertebrate muscle: correlations with anaerobic muscle work , 1984 .

[2]  K. Storey Phosphofructokinase from foot muscle of the whelk, Busycotypus canaliculatum: evidence for covalent modification of the enzyme during anaerobiosis. , 1984, Archives of biochemistry and biophysics.

[3]  E. Schaftingen Fructose 2,6-bisphosphate. , 2006 .

[4]  D. Livingstone,et al.  Carbohydrate metabolism of gastropods , 1983 .

[5]  P. Cortesi,et al.  Energy metabolism of bivalves at reduced oxygen tensions , 1992 .

[6]  K. Storey Suspended animation: the molecular basis of metabolic depression , 1988 .

[7]  A. Verhoeven,et al.  Anaerobic energy metabolism in isolated adductor muscle of the sea musselMytilus edulis L. , 1982, Journal of Comparative Physiology.

[8]  E. Helmerhorst,et al.  Microcentrifuge desalting: a rapid, quantitative method for desalting small amounts of protein. , 1980, Analytical biochemistry.

[9]  D. V. Slyke 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 , 1922 .

[10]  K. Storey,et al.  Tissue specific isozymes of pyruvate kinase in the channelled whelkBusycotypus canaliculatum: enzyme modification in response to environmental anoxia , 1985, Journal of Comparative Physiology □ B.

[11]  K. Storey Mechanisms of glycolytic control during facultative anaerobiosis in a marine mollusc: tissue-specific analysis of glycogen phosphorylase and fructose-2,6-bisphosphate , 1988 .

[12]  K. Storey,et al.  Phosphorylation in vivo of red-muscle pyruvate kinase from the channelled whelk, Busycotypus canaliculatum, in response to anoxic stress. , 1984, European journal of biochemistry.

[13]  J. Kluytmans,et al.  Anaerobic capacities and anaerobic energy production of some Mediterranean bivalves , 1983 .

[14]  K. Storey,et al.  Organ-specific regulation of phosphofructokinase during facultative anaerobiosis in the marine whelk Busycotypus canaliculatum , 1991 .

[15]  E. Schaftingen,et al.  Fructose 2,6-bisphosphate. , 1982, Biochemical Society transactions.

[16]  G. Somero,et al.  Buffering capacity of vertebrate muscle: Correlations with potentials for anaerobic function , 1981, Journal of comparative physiology.

[17]  F. Ghisotti,et al.  OSSERVAZIONI SULLA POPOLAZIONE DI SCAPHARCA, INSEDIATASI IN QUESTI ULTIMI ANNI SU UN TRATTO DEL LITORALE ROMAGNOLO 000 , 1976 .

[18]  K. Storey,et al.  Buffering Capacities of the Tissues of Marine Molluscs , 1984, Physiological Zoology.

[19]  B. Michaelidis,et al.  Modification of pyruvate kinase from the foot muscle of Patella caerulea (L) during anaerobiosis , 1988 .

[20]  P. Cortesi,et al.  Energy metabolism during anaerobiosis and recovery in the posterior adductor muscle of the bivalve Scapharca inaequivalvis (Bruguiere) , 1989 .

[21]  J. Kluytmans,et al.  Organ specific changes in energy metabolism due to anaerobiosis in the sea mussel Mytilus edulis (L.) , 1980 .