Binding of oxygen and carbon monoxide to arthropod hemocyanin: an allosteric analysis.
The binding of oxygen and carbon monoxide to hemocyanin from the mangrove crab Scylla serrata and the lobster Homarus americanus has been studied by thin-layer optical absorption and front face fluorescence techniques. Three types of experiments were performed on subunit and oligomeric preparations of each hemocyanin: oxygen binding, carbon monoxide binding, and oxygen-carbon monoxide competition studies. The results obtained from the subunit preparations of dissociated oligomers from both hemocyanins show that the binding site can be ligated by either one oxygen or one carbon monoxide. The binding results obtained with the oligomeric samples of hemocyanin from both species cannot be described by the two-state MWC model [Monod, J., Wyman, J., & Changeux, J. P. (1965) J. Mol. Biol. 12, 88-118] since the data from the three types of binding experiments cannot be fit with a single set of binding constants. The MWC model has been extended by including a third allosteric form, and an analysis based on the three-state model is able to fit the data from the three types of experiments with the same set of binding constants. The comparison of the oxygen to carbon monoxide affinity ratios (kO2/kCO) indicates that the structure around the binding site of subunits in the T form oligomer is similar to that of the free subunits. The oligomeric forms of both these hemocyanins bind carbon monoxide with a weak but definite positive cooperativity. An analysis of the affinity ratios for the T, S, and R forms suggests that the high affinity of the R form results from a specific interaction between oxygen and binding site.
D000494:Allosteric Regulation; D000818:Animals; D003386:Brachyura / Q000378:metabolism*; D002248:Carbon Monoxide / Q000378:metabolism*; D006433:Hemocyanins / Q000378:metabolism*; D007700:Kinetics; D008024:Ligands; D008433:Mathematics; D008121:Nephropidae / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*; D011485:Protein Binding; D013045:Species Specificity
D008024:Ligands; D002248:Carbon Monoxide; D006433:Hemocyanins; D010100:Oxygen