Resonance raman studies of hemoglobins M: evidence for iron-tyrosine charge-transfer interactions in the abnormal subunits of Hb M Boston and Hb M Iwate.

Resonance Raman spectra have been obtained for Hb M Boston [His-E7(58) alpha leads to Tyr], Hb M Iwate [His-F8-(87) alpha leads to Tyr], and Hb M Milwaukee [Val-E11(67) beta leads to Glu]. The abnormal alpha subunits of Hb M Boston and Hb M Iwate exhibited the porphyrin nu10 band at 1628 and 1627 cm-1, respectively, which indicates that the ferric alpha hemes are five-coordinated in both Hb M Boston and Hb M Iwate. In addition to the porphyrin bands, four extra polarized lines were observed at 1607, 1506, 1278, and 603 cm-1 for the alpha abnormal subunit of Hb M Boston and at 1605, 1506, 1310, and 589 cm-1 for that of Hb M Iwate. By comparison with the vibrational spectra of Fe-tyrosine proteins and Fe-phenolate complexes, the 1605-1607- and 1506-cm-1 lines are assigned to the phenolate ring vibrations of the heme-coordinated tyrosine, and the 1278-cm-1 line of Hb M Boston and the 1310-cm-1 line of Hb M Iwate are assigned to the phenolate CO stretching mode. We propose that the 603-cm-1 line of Hb M Boston and the 589-cm-1 line of Hb M Iwate arise from the Fe-O(tyrosine) stretching mode. These four Raman lines are intensity enhanced upon the excitation around 475-520 nm, probably due to the presence of a charge-transfer interaction between Fe and Tyr. The dissimilarity of the Fe-O and phenolate CO stretching frequencies between Hb M Boston and Hb M Iwate, despite the similarity of frequencies of their porphyrin and phenolate ring modes, suggests that the heme-phenolate bonding angles differ between Hb M Boston and Hb M Iwate although both adopt the five-coordinate form with Tyr as the only axial ligand. The resonance Raman spectra of oxy- and deoxy-Hb M Milwaukee showed no anomaly and can be accounted for by those of the equimolar mixtures of aquomet- and oxy- or deoxy-Hb A.