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Binding of Urate and Caffeine to Hemocyanin Analyzed by Isothermal Titration Calorimetry
Journal of Experimental Biology (2001)
  • Michael A Menze, University of Louisville
  • Nadja Hellmann, Johannes Gutenberg Universitat, Mainz
  • Heinz Decker, Johannes Gutenberg Universitat, Mainz
  • Manfred K. Grieshaber
Haemocyanin serves as an oxygen carrier in the haemolymph of decapod crustaceans. The oxygen-binding behaviour of the pigment is modulated by the two major anaerobic metabolites, L-lactate and urate. The binding of these two metabolites to haemocyanin has been investigated mainly indirectly by following the effectorinduced changes in the oxygen-binding properties of the respiratory pigment. Only a few direct investigations of effector binding, employing ultracentrifugation techniques and equilibrium dialysis, have been carried out. No evidence for cooperative binding for either effector was detected using these methods. However, isothermal titration calorimetry (ITC) offers a useful tool to gain additional insight into the binding of effectors to these highly allosterically regulated macromolecules. By applying the ITC method to the fully oxygenated dodecameric haemocyanin of the lobster Homarus vulgaris, cooperativity in binding has been found for the urate analogue caffeine but not for urate itself: using urate and the urate analogue caffeine as ligands, two conformations of the oxygenated pigment were detected.
  • haemocyanin,
  • isothermal titration calorimetry,
  • oxygen binding,
  • effector,
  • caffeine,
  • urate,
  • cooperativity,
  • crustacean,
  • lobster,
  • Homarus vulgaris
Publication Date
March 1, 2001
Publisher Statement
This research is copyright 2001 The Company of Biologists Limited.
Citation Information
Michael A Menze, Nadja Hellmann, Heinz Decker and Manfred K. Grieshaber. "Binding of Urate and Caffeine to Hemocyanin Analyzed by Isothermal Titration Calorimetry" Journal of Experimental Biology Vol. 204 Iss. 5 (2001)
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