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.

[1]  A. Spisni,et al.  17O NMR investigation of the hydration of L-alanine and L-proline in water/Me2SO mixtures. , 1986, Biochemical and biophysical research communications.

[2]  S. Gill,et al.  Thin-layer fluorescence cell for ligand binding studies. , 1983, Analytical biochemistry.

[3]  M. Brunori,et al.  A spectrophotometric method to determine the amount of CO bound to hemocyanin. , 1983, Analytical biochemistry.

[4]  N. Ellerton,et al.  Hemocyanin--a current perspective. , 1983, Progress in biophysics and molecular biology.

[5]  M. Brunori,et al.  Reaction of carbon monoxide with hemocyanin: stereochemical effects of a non-bridging ligand. , 1981, Journal of molecular biology.

[6]  T. T. Herskovits,et al.  Light-scattering investigation of the subunit dissociation of Homarus americanus hemocyanin. Effects of salts and ureas. , 1981, Biochemistry.

[7]  K. V. van Holde,et al.  Allosteric properties and the association equilibria of hemocyanin from Callianassa californiensis. , 1979, Journal of molecular biology.

[8]  J. H. Ploegman,et al.  Structure of Helix pomatia oxy-beta-hemocyanin and deoxy-beta-hemocyanin tubular polymers. , 1979, European journal of biochemistry.

[9]  B. Barisas,et al.  Thermodynamic analysis of carbon monoxide binding by hemoglobin trout I. , 1979, Biophysical chemistry.

[10]  S. Gill,et al.  Membrane-covered thin-layer optical cell for gas-reaction studies of hemoglobin. , 1978, Analytical biochemistry.

[11]  J. Bonaventura,et al.  Analysis of the effect of three different allosteric ligands on oxygen binding by hemocyanin of the shrimp, Penaeus setiferus. , 1978, Biochemistry.

[12]  T. B. Freedman,et al.  A resonance Raman study of the copper protein, hemocyanin. New evidence for the structure of the oxygen-binding site. , 1976, Journal of the American Chemical Society.

[13]  E. Vanbruggen,et al.  FUNCTIONAL PROPERTIES OF CHEMICALLY MODIFIED HEMOCYANIN - FIXATION OF HEMOCYANIN IN LOW AND HIGH OXYGEN-AFFINITY STATE BY REACTION WITH A BIFUNCTIONAL IMIDO ESTER , 1975 .

[14]  J. Bonaventura,et al.  Carbon monoxide binding by hemocyanins of Limulus polyphemus, Busycon carica, and Callinectes sapidus , 1974 .

[15]  K. V. van Holde,et al.  Oxygen binding by callianassa californiensis hemocyanin. , 1974, Biochemistry.

[16]  J. O. Alben,et al.  Structure of the carbon monoxide binding site of hemocyanins studied by Fourier transform infrared spectroscopy. , 1972, Biochemistry.

[17]  G. Kegeles,et al.  Subunit interactions of lobster hemocyanin. I. Ultracentrifuge studies. , 1971, Archives of biochemistry and biophysics.

[18]  J. O. Alben,et al.  Infrared studies of carbon monoxide bound to hemocyanin and to simple copper complexes , 1970 .

[19]  J. L. Larimer,et al.  Lobster Hemocyanin: Properties of the Minimum Functional Subunit and of Aggregates , 1966, Science.

[20]  J. Changeux,et al.  ON THE NATURE OF ALLOSTERIC TRANSITIONS: A PLAUSIBLE MODEL. , 1965, Journal of molecular biology.

[21]  D. Marquardt.  An Algorithm for Least-Squares Estimation of Nonlinear Parameters , 1963 .

[22]  R. W. Root.  THE COMBINATION OF CARBON MONOXIDE WITH HEMOCYANIN , 1934 .

[23]  A. C. Redfield,et al.  THE RESPIRATORY PROTEINS OF THE BLOOD II. THE COMBINING RATIO OF OXYGEN AND COPPER IN SOME BLOODS CONTAINING HEMOCYANIN , 1928 .