Allosteric models for multimeric proteins: oxygen-linked effector binding in hemocyanin.
In many crustaceans, changing concentrations of several low molecular weight compounds modulates hemocyanin oxygen binding, resulting in lower or higher oxygen affinities of the pigment. The nonphysiological effector caffeine and the physiological modulator urate, the latter accumulating in the hemolymph of the lobster Homarus vulgaris during hypoxia, increase hemocyanin oxygen affinity and decrease cooperativity of oxygen binding. To derive a model that describes the mechanism of allosteric interaction between hemocyanin and oxygen in the presence of urate or caffeine, studies of oxygen, urate, and caffeine binding to hemocyanin were performed. Exposure of lobster hemocyanin to various pH values between 7.25 and 8.15 resulted in a decrease of p50. In this pH interval, p50 decreases from 95 to 11 Torr without effectors and from 49 to 6 Torr and from 34 to 5 Torr in the presence of 1 mM urate or caffeine, respectively. Thus, the allosteric effects induced by protons and urate or caffeine are coupled. In contrast, isothermal titration calorimetry did not reveal any differences in binding enthalpy (DeltaH degrees ) for urate or caffeine under either normoxic or hypoxic conditions at different pH values. Despite these apparently conflicting results, they can be explained by the nested MWC model if two different types of modulator binding sites are assumed, an allosteric and a nonallosteric type of site. Simulations of in vivo conditions with this model indicate that the naturally occurring modulator urate is physiologically relevant in H. vulgaris only during hypoxic conditions, i.e., either during environmental oxygen limitation or extensive exercise.
Menze
Michael A
MA
Institut für Zoophysiologie, Heinrich-Heine Universität, Universitätsstrasse 1, 40225 Düsseldorf, Germany. menze@lsu.edu
Hellmann
Nadja
N
Decker
Heinz
H
Grieshaber
Manfred K
MK
D000494:Allosteric Regulation; D000818:Animals; D001665:Binding Sites; D002110:Caffeine / Q000378:metabolism; D002151:Calorimetry; D006433:Hemocyanins / Q000737:chemistry / Q000378:metabolism*; D000860:Hypoxia / Q000378:metabolism; D008956:Models, Chemical; D008121:Nephropidae; D010100:Oxygen / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D014527:Uric Acid / Q000737:chemistry / Q000378:metabolism
D014527:Uric Acid; D002110:Caffeine; D006433:Hemocyanins; D010100:Oxygen