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The potentiating effect of purine bases and some of their derivatives on the oxygen affinity of haemocyanin from the crayfishAustropotamobius pallipes


  1. 1.

    At pH 7.8 theP 50 of dialysedAustropotamobius pallipes haemolymph (P 50=1.7 Torr) was 2.9 Torr greater than that of nondialysed haemolymph (P 50=4.6 Torr). This difference was not attributable to the specific effect ofl-lactate but rather to unidentified factors (U.F.) which increase the affinity of haemocyanin for oxygen.

  2. 2.

    The presence of 0.35 mM urate in the haemolymph of this species was confirmed as was the potentiating effect of this ion on oxygen affinity. In this study 2 mM urate reduced theP 50 of dialysed haemolymph by approximately 50% from 4.6 Torr to 2.1 Toor.

  3. 3.

    The specificity of the haemocyanin for a specific purine structure was demonstrated to be low as the urate analogues, caffeine and theobromine, isolated from plants also increased the oxygen affinity of dialysed haemolymph.

  4. 4.

    Metabolites arising from the degradation of ADP/ATP to urate can also increase haemocyanin oxygen affinity to a variable extent but not as effectively as urate. Compounds in which the purine ring was cleaved evoked very little or no specific enhancement of haemocyanin oxygen affinity.

  5. 5.

    A combination of purine bases and derivatives, shown to produce individually an increase in oxygen affinity, did not produce an effect in excess of that due to urate alone. The presence of high concentrations of urate in the haemolymph abolishes the specific effect ofl-lactate.

  6. 6.

    Additive and synergistic effects ofl-lactate and the various purine derivatives may account for 80% of the U.F. effect.

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  1. Atkinson DE (1968) The energy charge of the adenylate pool as a regulating parameter. Interaction with feedback modifiers. Biochemistry 7:4030–4034

  2. Atkinson DE (1970) Enzymes as control elements in metabolite regulation. In: Boyer PD (ed) The Enzymes. 3rd edition. Academic Press, New York

  3. Atkinson DE, Camien MN (1982) The role of urea synthesis in the removal of metabolic bicarbonate and the regulation of blood pH. Curr Topics Cell Reg 21:261–301

  4. Benesch RE, Benesch R (1967) The effect of organic phosphates from the human crythrocyte on the allosteric properties of hemoglobin. Biochem Biophys Res Com 26:162–167

  5. Binns R (1969) The physiology of the antennal gland ofCarcinus maenas. V. Some nitrogenous constituents in the blood and urine. J Exp Biol 51:41–45

  6. Booth CE, McMahon BR, Pinder AW (1982) Oxygen uptake and the potentiating effects of increased hemolymph lactate on oxygen transport during exercise in the blue crab,Callinectes sapidus. J Comp Physiol 148:111–121

  7. Bridges CR, Bicudo JEPW, Lykkeboe G (1979) Oxygen content measurements in blood containing haemocyanin. Comp Biochem Physiol 62A:457–462

  8. Bridges CR, Morris S, Grieshaber MK (1984) Modulation of haemocyanin oxygen affinity in the intertidal prawnPalaemon elegans (Rathke). Resp Physiol 57:189–200

  9. Chanutin A, Curnish RR (1967) Effect of organic and inorganic phosphates on the oxygen equilibrium of human erythrocytes. Archs Biochem Biophys 121:96–102

  10. Claybrook DL (1983) Nitrogen metabolism. In: Mantel LH (ed) The biology of Crustacea, Vol 5. Academic Press, New York, pp 163–213

  11. Clegg JS, Warner AH, Finamore FJ (1967) Evidence for the function of P1, P4-diguanosine 5′-tetraphosphate in the development ofArtemia salina. J Biol Chem 242:1938–1943

  12. Delaunay H (1931) L'excretion azotée des invertébrés. Biol Rev Cambridge Philos Soc 6:265–301

  13. Dresel EIB, Moyle V (1950) Nitrogenous excretion in amphipods and isopods. J Exp Biol 27:210–225

  14. Engel PC, Jones JB (1978) Causes and elimination of erratic blanks in enzymatic metabolite assays involving the use of NAD+ in alkaline hydrazine buffers: improved conditions for the assay ofl-glutamate,l-lactate, and other metabolites. Anal Biochem 88:475–484

  15. Gifford CA (1968) Accumulation of uric acid in the land crab,Cardisoma guanhumi. Am Zool 8:521–528

  16. Graham RA, Mangum CP, Terwilliger RC, Terwilliger N (1983) The effect of organic acids on oxygen binding of hemocyanin from the crabCancer magister. Comp Biochem Physiol 74A:45–50

  17. Greenaway P (1974) Total body calcium and haemolymph calcium concentrations in the crayfishAustropotamobius pallipes (Lereboullet). J Exp Biol 61:19–26

  18. Gutmann I, Wahlfeld AW (1974) L-(+)-lactate determined with lactate dehydrogenase and NAD. In: Bergmeyer HU (ed) Methods in enzymatic analysis, 2nd edn. Verlag Chemie, Weinheim and Academic Press Inc, London New York

  19. Harris RR, Chantler EN, Bannister WH (1975) The oxygen affinity ofCarcinus maenas haemocyanin — A calcium insensitive, dialysis induced decrease in O2 affinity. Comp Biochem Physiol 52A:189–191

  20. Henry RP, Cameron JN (1981) A survey of blood and tissue nitrogen compounds in terrestrial decapods in Palau. J Exp Zool 218:83–88

  21. Hernandorena A (1979) Relationship between purine and pyrimidine dietary requirements andArtemia morphogenesis. Comp Biochem Physiol 62B:7–12

  22. Hogben LJ (1926) Some observations on the dissociation of haemocyanin by the colormetric method. Brit J Exp Biol 3:225–238

  23. Isaaks RE, Harkness DR (1980) Erythrocyte organic phosphates and hemoglobin function in birds, reptiles and fishes. Am Zool 20:115–129

  24. Johnson BA, Bonaventura C, Bonaventura J (1984) Allosteric modulation ofCallinectes sapidus haemocyanin by binding ofl-lactate. Biochemistry 23:872–878

  25. Jokumsen A, Weber RE (1980) Hemoglobin oxygen binding properties in the blood ofXenopus laevis; influences of estivation, salinity and temperature-acclimation. J Exp Biol 86:19–37

  26. Larimer JL, Riggs AF (1964) Properties of hemocyanins — I. The effect of calcium ions on the oxygen equilibrium of crayfish haemocyanin. Comp Biochem Physiol 13:35–46

  27. Lehninger AL (1976) Biochemistry, the molecular basis of cell structure and function, 2nd edn. New York, Worth Publishers, p 1104

  28. Mangum CP (1983a) Adaptability and inadaptability among HcO2 transport systems: an apparent paradox. In: Wood EJ (ed) Structure and function of invertebrate respiratory proteins. Life Chem Repts Supp 1. Harwood Academic Publishers, Chur London New York, pp 333–352

  29. Mangum CP (1983b) On the distribution of lactate sensitivity among hemocyanins. Mar Biol Lett 4:139–149

  30. Morris S, Bridges CR (1985) An investigation of haemocyanin oxygen affinity in the semi-terrestrial crabOcypode saratan Forsk. J Exp Biol 117:(in press)

  31. Morris S, Bridges CR, Grieshaber MK (1985a) Evidence for the presence and specificity of an unidentified plasma cofactor increasing haemocyanin oxygen affinity in two natantid crustaceans. J Exp Zool 234:151–155

  32. Morris S, Bridges CR, Grieshaber MK (1985b) A new role for uric acid: Modulator of haemocyanin oxygen affinity in crustaceans. J Exp Zool 235:135–139

  33. Newsholme EA, Start C (1977) Regulation in metabolism. John Wiley, London New York Sydney Toronto, p 349

  34. Nickerson KW, van Holde KFE (1971) A comparison of molluscan and arthropod hemocyanin-I. Circular dichroism and adsorption spectra. Comp Biochem Physiol 39B:855–872

  35. Rossi-Fanelli A, Antonini E, Caputo A (1964) Hemoglobin and myoglobin. Adv Protein Chem 19:74–222

  36. Sick H, Gersonde K (1969) Method for continuous registration of O2-binding curves of hamoproteins by means of a diffusion chamber. Analyt Biochem 32:362–376

  37. Stedman E, Stedman E (1926) The influence of hydrogen ion concentration in the dissociation curve of oxyhaemocyanin from the edible crab (Cancer pagurus). Biochem J 20:949–956

  38. Taylor AC, Morris S, Bridges CR (1985) Oxygen and carbon dioxide transporting properties of the blood of three sublittoral species of burrowing crab. J Comp Physiol 155:733–742

  39. Taylor EW, Wheatly MG (1980) Ventilation, heart-rate and respiratory gas exchange in the crayfishAustropotamobius pallipes (Lereboullet) submerged in normoxic water and after 3 h exposure in air at 15°C. J Comp Physiol 138:67–78

  40. Taylor EW, Wheatly MG (1981) The effect of long-term aerial exposure on heart rate, ventilation, respiratory gas exchange and acid-base status in the crayfishAustropotamobius pallipes. J Exp Biol 92:109–124

  41. Tetens V, Wells RMG (1984) Oxygen binding properties of the blood and hemoglobin solutions in the carpet shark (Cephaloscyllium isabella): Role of ATP and urea. Comp Biochem Physiol 79A:165–168

  42. Truchot J-P (1971) Etude comparée de la fixation de l'oxygène par le sérum de cinq espèces de Crustacés Decapodes Brachyoures. CR Acad Soc Paris 272:2706–2709

  43. Truchot J-P (1975) Factors controlling the in vitro and in vivo oxygen affinity of the haemocyanin in the crabCarcinus maenas (L.). Respir Physiol 24:173–189

  44. Truchot J-P (1980) Lactate increases the oxygen affinity of crab haemocyanin. J Exp Zool 214:205–208

  45. Van Denbos G, Finamore FJ (1974) An unusual pathway for the synthesis of ATP by the purine-requiring organismArtemia salina. J Biol Chem 249:2816–2818

  46. Weber RE, Wells RMG, Rossetti JE (1983) Allosteric interactions governing oxygen equilibria in the haemoglobin systems of the spiny dogfish,Squalus acanthias. J Exp Biol 103:109–120

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Correspondence to S. Morris.

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Morris, S., Bridges, C.R. & Grieshaber, M.K. The potentiating effect of purine bases and some of their derivatives on the oxygen affinity of haemocyanin from the crayfishAustropotamobius pallipes . J Comp Physiol B 156, 431–440 (1986). https://doi.org/10.1007/BF01101106

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  • Oxygen
  • Caffeine
  • Human Physiology
  • Synergistic Effect
  • Purine