Amphitrite ornata erythrocruorin. II. Molecular controls of function.
In the marine terebellid worm Amphitrite ornata the vascular fluid contains a high molecular weight erythrocruorin, while cells of the coelom contain a monomeric hemoglobin. The structural integrity of the erythrocruorin molecule is known to be dependent on the presence of a minimal concentration of divalent cations (1-3 mM) in the medium. The functional properties of Amphitrite erythrocruorin are also affected by cations. The oxygen affinity tends to increase with increasing cation concentration and the degree of cooperative interactions, expressed in the kinetics and equilibria of ligand binding, goes through a maximum. Maximal Hill coefficients of 3-4 are observed with 50 mM CaCl2, 50 mM MgCl2 or 1 M NaCl in measurements at the physiological pH of 7.75. Only 2 mM CaCl2 is required for maximal cooperativity at pH 8.5. This suggests partial deprotonation of the cation binding site at high pH. It is somewhat unusual that pH effects on cooperativity are reversible, since this is not a common feature of the giant erythrocruorin molecules. The oxygen binding experiments revealed a marked effect of divalent cations of Amphitrite erythrocruorin at high pH and cation concentration. Above pH 8.5, at 50 mM CaCl2 and 12 degrees C, the erythrocruorin will form a polymer upon deoxygenation. This polymerization is readily reversible by bringing the temperature for 12 to 20 degrees C or by oxygenation. Under physiological conditions of pH and cation concentration and at 12 degrees C, the erythrocruorin and the monomeric coelomic hemoglobin require a similar oxygen pressure for half saturation. However, the allosteric regulation of function is absent for the coelomic protein.
D000818:Animals; D002122:Calcium Chloride / Q000494:pharmacology; D002248:Carbon Monoxide / Q000378:metabolism; D002413:Cations, Divalent; D004902:Erythrocruorins / Q000378:metabolism*; D006454:Hemoglobins / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D046911:Macromolecular Substances; D008274:Magnesium / Q000494:pharmacology; D015636:Magnesium Chloride; D010100:Oxygen / Q000378:metabolism; D011077:Polychaeta / Q000032:analysis*; D012965:Sodium Chloride / Q000494:pharmacology
D002413:Cations, Divalent; D004902:Erythrocruorins; D006454:Hemoglobins; D046911:Macromolecular Substances; D015636:Magnesium Chloride; D012965:Sodium Chloride; D002248:Carbon Monoxide; D008274:Magnesium; D002122:Calcium Chloride; D010100:Oxygen