Kinetic properties and electron paramagnetic resonance spectra of the nitric oxide derivative of hemoglobin components of trout (Salmo irideus).
The binding of nitric oxide to hemoglobin components of trout (Salmo irideus), i.e., Hb trout I and Hb trout IV, has been studied by optical and electron paramagnetic resonance spectroscopy. Kinetic studies show that the Root effect in Hb trout IV is operative also for NO, since a large increase in the dissociation velocity constant (j(4)) is observed as the pH is decreased below 7. Moreover, the time course of the displacement of NO by CO is heterogeneous, suggesting that alpha and beta chains may have different j(4) values. Low-temperature X-band electron paramagnetic resonance spectra have been recorded with Hb trout I and IV saturated with NO at different pH values. The spectra of Hb trout IV are strongly pH-dependent. The high-pH form (pH 8.1) shows axial symmetry and no resolved hyperfine splitting, while the low-pH form is rhombic with a hyperfine splitting of 6.5 G in the g(z) region. The latter form reflects a more distorted site with a more significant delocalization of the unpaired electron on the proximal histidine; both features indicate a destabilization of the ligand binding at low pH. On the other hand, spectra of Hb trout I are axial at both pH values, with hyperfine splitting of 16.5 G, indicating that the site is not distorted and interacts with the ligand very strongly at either pH.
D000818:Animals; D004578:Electron Spin Resonance Spectroscopy; D006454:Hemoglobins / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008024:Ligands; D009569:Nitric Oxide / Q000097:blood; D011485:Protein Binding; D012487:Salmonidae / Q000378:metabolism*
D006454:Hemoglobins; D008024:Ligands; D009569:Nitric Oxide