Differences in iron-fluoride bonding between the isolated subunits of human methemoglobin fluoride and sperm whale metmyoglobin fluoride as measured by resonance Raman spectroscopy.

The heme geometries of the isolated alpha and beta subunits of human methemoglobin fluoride (HbIIIF) and sperm whale metmyoglobin fluoride (MbIIIF) have been examined by exciting their Raman spectra within their ca. 6000-A charge-transfer absorption bands. The Fe-F stretching vibration at 471 cm-1 in the beta subunits shifts to 466 cm-1 in the alpha subunits and to 461 cm-1 in MbIIIF. The Fe-F bond is estimated to elongate by 0.02 A in the alpha subunits and 0.03 AZ in MbIIIF compared with that in the beta subunits. This bond elongation is interpreted to result from an increased iron displacement toward the proximal histidine side of the heme in the series MbIIIF greater than alpha greater than beta. A comparison of the isolated subunit spectra with that of tetrameric HbIIIF indicates little change occurs in isolated subunit heme geometry upon association into tetrameric HbIIIF. A correlation is found between the gamma max of the 600-A charge-transfer absorption band and the Fe-F bond length. Elongation of the Fe-F bond is associated with a shift of the absorption spectral maximum to a longer wavelength. However, the absorption spectral shift induced by the inositsol hexaphosphate induced R leads to T conversion does not result from a change in the Fe-F stretching frequency (+/- 0.5 cm-1). In contrast, frequency shifts are observed for heme macrocyclic vibrational modes. The data are interpreted to indicate that the effect of the R leads to T conversion in HbIIIF is to perturb heme macrocycle conformation without altering the heme out-of-plane iron distance or the Fe-F bond length.