Advertisement

Journal of Comparative Physiology B

, Volume 155, Issue 6, pp 723–731 | Cite as

Isolation and characterisation of tissue-specific isoenzymes of D-lactate dehydrogenase from muscle and hepatopancreas ofLimulus polyphemus

  • Karl-Heinz Carlsson
  • Gerd Gäde
Article

Summary

  1. 1.

    Isoenzymes of dimeric,d-lactate specific lactate dehydrogenase from hepatopancreas and muscle tissue of the horseshoe crab,Limulus polyphemus, have been purified to homogeneity by chromatographic techniques.

     
  2. 2.

    Separation of the three muscle isoenzymes, designated CC, CD and DD, was achieved by ion-exchange chromatography, whereas the three hepatopancreas isoenzymes, designated AA, AB and BB, were separated by chromatofocusing.

     
  3. 3.

    Kinetic studies suggest that hepatopancreasd-LDH is analogous to vertebrate heart-typel-LDH; whereas horseshoe crab muscled-LDH resembles vertebrate M-typel-LDH; for example, hepatopancreasd-LDH is strongly substrate inhibited by pyruvate, whereas muscled-LDH is not. Moreover, hepatopancreasd-LDH has a higher affinity ford-lactate and NAD+ than muscled-LDH. Finally, at physiological pH values, it appears that the hepatopancreas isoenzymes are strongly poised in the direction ofd-lactate oxidation.

     
  4. 4.

    The kinetic data indicate the potential for organ partitioning ofd-lactate formation andd-lactate oxidation:d-lactate produced in the muscles may be transported via the hemolymph to the hepatopancreas for oxidation.

     

Keywords

Lactate Pyruvate Human Physiology Muscle Tissue Lactate Dehydrogenase 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Beaumont AR, Day TR, Gäde G (1980) Genetic variations at the octopine dehydrogenase locus in the adductor muscle ofCerastoderma edule (L.) and six other bivalve species. Mar Biol Lett 1:137–148Google Scholar
  2. Bradford MM (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analyt Biochem 72:248–254Google Scholar
  3. Carlsson K-H, Gäde G (1981) Anaerobic metabolism of the common cockle,Cardium edule. III. Purification and physiological role of D-lactate dehydrogenase from muscle and gill tissue. Comp Biochem Physiol 69B:715–724Google Scholar
  4. Dunbar CO (1923) Kansas Permian insects. Part 2. Paleolimulus, a new genus of Paleozoic Xiphosura with notes on other genera. Am J Sci Ser 5:443–454Google Scholar
  5. Ellington WR (1983a) The extent of intracellular acidification during anoxia in the catch muscles of two bivalve molluscs. J Exp Zool 227:313–317Google Scholar
  6. Ellington WR (1983b) Phosphorus nuclear magnetic resonance studies of energy metabolism in molluscan tissues: Effect of anoxia and ischemia on the intracellular pH and high energy phosphates in the ventricle of the whelk,Busycon contrarium. J Comp Physiol 153:159–166Google Scholar
  7. Everse J, Kaplan NO (1973) Lactate dehydrogenase: structure and function. Adv Enzymol 37:61–148Google Scholar
  8. Falkowski PG (1974) Facultative anaerobiosis inLimulus polyphemus: phosphoenolpyruvate carboxykinase and heart activities. Comp Biochem Physiol 49B:749–759Google Scholar
  9. Gäde G (1980a) A comparative study of octopine dehydrogenase isoenzymes in gastropod, bivalve and cephalopod molluscs. Comp Biochem Physiol 76B:73–78Google Scholar
  10. Gäde G (1980b) Biological role of octopine formation in marine molluscs. Mar Biol Lett 1:121–135Google Scholar
  11. Gäde G, Carlsson K-H (1984) Purification and characterisation of octopine dehydrogenase from the marine nemerteanCerebratulus lacteus (Anopla: Heteronemertea): comparison with the scallop octopine dehydrogenase. Mar Biol 79: 39–45Google Scholar
  12. Gäde G, Grieshaber M (1976) Anaerobic metabolism of the common cockle,Cardium edule. II. Partial purification and properties of lactate dehydrogenase and octopine dehydrogenase. A comparative study. Arch Int Physiol Biochim 84:735–752Google Scholar
  13. Long GL (1976) The stereospecific distribution and evolutionary significance of invertebrate lactate dehydrogenase. Comp Biochem Physiol 55B:77–83Google Scholar
  14. Long GL, Kaplan NO (1968) D-lactate specific pyridine nucleotide lactate dehydrogenase in animals. Science 162:685–686Google Scholar
  15. Long GL, Kaplan NO (1973a) Diphosphopyridine nucleotide-linked D-lactate dehydrogenase from horseshoe crab,Limulus polyphemus and the seaworm,Nereis virens. I. Physical and chemical properties. Arch Biochem Biophys 154:696–710Google Scholar
  16. Long GL, Kaplan NO (1973b) Diphosphopyridine nucleotide-linked D-lactate dehydrogenase from horseshoe crab,Limulus polyphemus, and the seaworm,Nereis virens. II. Catalytic properties. Arch Biochem Biophys 154:711–725Google Scholar
  17. Long GL, Ellington WR, Duda TF (1979) Comparative enzymology and physiological role of D-lactate dehydrogenase from foot muscle of two gastropod molluscs. J Exp Zool 207:237–248Google Scholar
  18. Plaxton WC, Storey KB (1982) Tissue specific isozymes of alanopine dehydrogenase in the channeled whelkBusycotypus canaliculatum. Can J Zool 60:1568–1572Google Scholar
  19. Pörtner HO, Grieshaber MK, Heisler N (1984) Anaerobiosis and acid-base status in marine invertebrates: effect of environmental hypoxia on extracellular and intracellular pH inSipunculus nudus L. J Comp Physiol B 155:13–20Google Scholar
  20. Storey KB, Storey JM (1979) Kinetic characterization of tissue-specific isoenzymes of octopine dehydrogenase from mantle muscle and brain ofSepia officinalis. Eur J Biochem 93:545–552Google Scholar
  21. Tuengler P, Stein TN, Long GL (1980) Studies on the active center of D- and L-lactate dehyhrogenases using oxamatediaminohexyl-Sepharose affinity chromatography. Proc Natl Acad Sci USA 77:5832–5836Google Scholar

Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • Karl-Heinz Carlsson
    • 1
  • Gerd Gäde
    • 2
  1. 1.Institut für Pharmakologie und ToxikologieUniversität des SaarlandesHomburg/SaarFederal Republic of Germany
  2. 2.Institut für Zoologie IV der Universität DüsseldorfDüsseldorf 1Federal Republic of Germany

Personalised recommendations