On the interaction of small molecules with hemoglobin: orientational effects of hydration layers in protein crystal.
The spin label TEMPO does not show a binding to hemoglobin molecule in solution. In the crystal however the spin label is bound and a considerable anisotropy in the ESR spectra is observed similar to that with covalent spin labeling. Since TEMPO is a small spherical molecule the anisotropy should be a consequence of the ordering in the crystal packing observed for the hydration layers. Besides the anisotropic sites (one per asymmetric unit) an isotropic signal is apparent. The population of these sites is sensitive to the temperature and above around 30 degrees C a transition is observed of the label from the anisotropic site to the isotropic one. This is consistent with a change in the hydration structure above this temperature so that the spin label is sensitive to the reorganization of water or crystallization. Results of simulations based on the relaxation theory in liquids are compared for different hemoglobin physical states: crystal, powder and solution. It is shown that the ESR parameters obtained in the crystal are very different from those used for spectral stimulation at low temperature.
D003460:Crystallization; D004578:Electron Spin Resonance Spectroscopy; D006454:Hemoglobins; D006801:Humans; D011208:Powders; D012996:Solutions; D013113:Spin Labels; D013696:Temperature
D006454:Hemoglobins; D011208:Powders; D012996:Solutions; D013113:Spin Labels