Structural states and transitions of carp hemoglobin.
The wide ligand affinity range previously observed for carp hemoglobin is bounded at both extremes by regions of constant affinity. Within these regions, pH, organic phosphates, and the extent of ligand binding have no effect on the measured affinity and the cooperativity of ligand binding is greatly reduced or absent. The rates of CO recombination to fully and partially unliganded carp hemoglobin, under various organic phosphate and pH conditions, are shown to reflect this behavior. Constant kinetic rates are seen to directly correspond to the regions of constant affinity. Therefore, these are taken to be single protein conformations, one of high and one of low ligand affinity. In the simplest view, these conformations represent the R and T states of a two-state model, and most of the properties of carp hemoglobin are explained quite well within this framework. Increases in either hydrogen or phosphate ion concentrations favor the stabilization of the low affinity structure of even fully liganded carp hemoglobin. We have studied the structural transition from high to low affinity by monitoring the absorption spectra of carp hemoglobins at constant pH as a function of organic phosphate concentration. We find that different spectra are induced in both carp methemoglobin and cyanomethemoglobin by inositol hexaphosphate addition. Furthermore, the dependence of the magnitude of the spectral changes on pH and organic phosphate concentration is the close agreement with that predicted from studies of the ligand binding properties of the molecule.
D000818:Animals; D002248:Carbon Monoxide / Q000097:blood; D002347:Carps / Q000097:blood*; D003530:Cyprinidae / Q000097:blood*; D006454:Hemoglobins; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008706:Methemoglobin; D010777:Photochemistry; D010833:Phytic Acid / Q000097:blood; D011487:Protein Conformation; D013053:Spectrophotometry
D006454:Hemoglobins; D010833:Phytic Acid; D002248:Carbon Monoxide; D008706:Methemoglobin