Summary
The changes in some kinetic properties of pyruvate kinase from the mantle muscle and the hepatopancreas of Patella caerulea, over a time-course of 16 or 8 h of acute hypoxia were studied. Up to the second hour of anaerobiosis PK exhibited changes of the kinetic properties in the direction of activation, whereas in the late stages of anaerobiosis in both tissues examined these changes occurred in the direction of inactivation. Although PKs from these tissues seemed to be regulated in a similar pattern during the early stages of anaerobiosis, they exhibited different kinetic properties under both normoxic and hypoxic conditions. Two forms of PK were found to be present in both the mantle muscle and the hepatopancreas of P. caerulea with different kinetic properties. The ratio of the enzyme units distributed between the two forms of PK (designated peak I and peak II) was varied during anaerobiosis indicating that the two forms of PK are interconvertible.
Article PDF
Similar content being viewed by others
Abbreviations
- ADP :
-
adenosine diphosphate
- ATP :
-
adenosine triphosphate
- EGTA :
-
ethyleneglycol-bis-β-aminoethylether N,N,N′,N′-tetraacetic acid
- PK :
-
pyruvate kinase
- PEP :
-
phosphoenolpyruvate
- PMSF :
-
phenylmethysulfonyl fluoride
References
Bucher T, Pfleiderer G (1955) In: Methods in enzymology, Vol. 1, Colowick SP, Kaplan NO (eds) Academic Press, New York, pp 435–440
De Vooys CGN, Holwerda DA (1986) Anaerobic metabolism in sublittoral living Mytilus galloprovincialis Lam. in the Mediterranean. III. The effect of anoxia and osmotic stress on some kinetic parameters of adductor muscle pyruvate kinase. Comp Biochem Physiol 85:217–221
Ebberink RHM, De Zwaan A (1980) Control of glycolysis in the posterior adductor muscle of the sea mussel Mytilus edulis. J Comp Physiol 137:165–171
Giles IG, Poat PC, Munday KA (1976) Purification and properties of puruvate kinase from the hepatopancreas of Carcinus maenas. Biochem J 153:127–134
Hochachka PW (1986) Defence strategies against hypoxia and hypothermia. Science 231:234–241
Hochachka PW, Somero GN (1984) Biochemical adaptation. Princeton University Press, Princeton, NJ
Holwerda DA, De Zwaan A (1973) Kinetic and molecular characteristics of allosteric pyruvate kinase from muscle tissue of the sea mussel Mytilus edulis L. Biochim Biophys Acta 309:296–306
Holwerda DA, Kruitwagen ECJ, De Bont AMTH (1981) Regulation of pyruvate kinase and phosphoenolpyruvate carboxykinase activity during anaerobiosis in Mytilus edulis (L). Mol Physiol 1:165–171
Holwerda DA, Veenhof PR, Van Heugten HAA, Zandee DI (1983) Modification of mussel pyruvate kinase during anaerobiosis and after temperature acclimation. Mol Physiol 3:225–234
Holwerda DA, Veenhof PR, de Zwaan A (1984) Physiological and biochemical investigations of the ecological relevance of anaerobiosis in bivalves. I. The changes in activity of mussel adductor muscle and mantle pyruvate kinase during aerial exposure and reimmersion. Mar Biol Lett 5:185–190
Job D, Crochet C, Dhien A, Chambaz EM (1978) A rapid method for screening inhibitor effects: Determination of I50 and its standard deviation. Anal Biochem 84:68–77
Leon O, Moran A, Gonzalez R (1982) Purification and characterization of pyruvate kinase from muscle of the sea mollusc Concholepas concholepas. Comp Biochem Physiol 72:65–69
Michaelidis B (1984) Study of energy metabolism in the foot of the sea mollusc P. caerulea (L). Thesis, University of Thessaloniki, Thessaloniki Greece
Michaelidis B, Beis I (1990) Studies on the anaerobic energy metabolism in the foot muscle of marine gastropod Patella caerulea (L). Comp Biochem Physiol 95B:493–500
Michaelidis B, Lazou A, Beis I (1985) Purification, catalytic and regulatory properties of pyruvate kinase from the foot of P. caerulea (L). Comp Biochem Physiol 82:405–412
Michaelidis B, Gaitanaki C, Beis I (1988) Modification of pyruvate kinase from the foot muscle of P. caerulea (L) during anaerobiosis. J Exp Zool 248:264–271
Moran A, Gonzalez R, Leon O (1983) Studies of the effects of phenylalanine and pH on pyruvate kinase from the muscle of the sea mollusc Concholepas concholepas. Comp Biochem Physiol 75:603–605
Plaxton WC, Storey KB (1984a). Purification and properties of anaerobic and anoxic forms of pyruvate kinase from red muscle of the channelled whelk, Busycotypus canaliculatum. Eur J Biochem 143:257–265
Plaxton WC, Storey KB (1984b) Phosphorylation in vivo of red muscle pyruvate kinase from the channelled whelk, Busycotypus canaliculatum: Enzyme modification in response to anoxic stress. Eur J Biochem 143:267–272
Plaxton WC, Storey KB (1985a) Tissue specific isozymes of pyruvate kinase in the channelled whelk, Busycotypus canaliculatum: Enzyme modification in response to anoxia. J Comp Physiol 155:291–296
Plaxton WC, Storey KB (1985b) Purification and properies of aerobic and anoxic forms of pyruvate kinase from the hepatopancreas of a channelled whelk Busycotypus canaliculatum. Arch Biochem Biophys 243:195–205
Sanchez JL, Abad M, Garcia Martin LO, Galanza A (1984) Purification and properties of pyruvate kinase from the hepatopancreas of Mytilus galloprovincialis (L) Comp Biochem Physiol 78B:873–879
Shick JM, De Zwaan A, De Bont (1983) Anoxic metabolic rate in the mussel Mytilus edulis (L) estimated by simultaneous direct calorimetry and biochemical analysis. Physiol Zool 56:56–63
Storey KB (1985) A re-evaluation of the Pasteur effect: new mechanisms in anaerobic metabolism. Mol Physiol 8:439–461
Zammit VA, Newsholme EA (1978) Properties of pyruvate kinase and phosphoenolpyruvate carboxykinase in relation to the direction and regulation of phosphoenolpyruvate metabolism in muscles of the frog and marine invertebrates. Biochem J 174:979–987
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Gaitanaki, C., Papadopoulos, A. & Beis, I. Time-course of covalent modification of pyruvate kinase during anaerobiosis in the mantle muscle and the hepatopancreas of the limpet Patella caerulea (L). J Comp Physiol B 160, 529–535 (1990). https://doi.org/10.1007/BF00258980
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00258980