UV resonance Raman studies of alpha-nitrosyl hemoglobin derivatives: Relation between the alpha 1-beta 2 subunit interface interactions and the Fe-histidine bonding of alpha heme

Human alpha-nitrosyl beta-deoxy hemoglobin A, alpha(NO)beta(deoxy), is cons idered to have a T (tense) structure with the low O-2 affinity extreme and the Fe-histidine (His87) (Fe-His) bond of alpha heme cleaved. The Fe-His bo nding of alpha heme and the intersubunit interactions at the alpha 1-beta 2 contact of alpha(NO)-Hbs have been examined under various conditions with EPR and UV resonance Raman (UVRR) spectra excited at 235 nm, respectively. NOHb at pH 6.7 gave the UVRR spectrum of the R structure, but in the presen ce of inositol-hexakis-phosphate (IHP) for which the Fe-His bond of the alp ha heme is broken, UVRR bands of Trp residues behaved half-T-like while Tyr bands remained R-like. The half-ligated nitrosylHb, alpha(NO)beta(deoxy), in the presence of IHP at pH 5.6, gave T-like UVRR spectra for both Tyr and Trp, but binding of CO to its beta heme (alpha(NO)beta(CO)) changed the UV RR spectrum to half-T-like. Binding of NO to its beta heme (NOHb) changed t he UVRR spectrum to 70% T-type for Trp but almost R-type for Tyr. When the pH was raised to 8.2 in the presence of IHP, the UVRR spectrum of NOHb was the same as that of COHb. EPR spectra of these Hbs indicated that the Fe-Hi s bond of alpha(NO) heme is partially cleaved. On the other hand, the UVRR spectra of alpha(NO)beta(deoxy) in the absence of IHP at pH 8.8 showed the T-like UVRR spectrum, but the EPR spectrum indicated that 40-50% of the Fe- His bond of alpha hemes was intact. Therefore, it became evident that there is a qualitative correlation between the cleavage of the Fe-His bond of al pha heme and T-like contact of Trp-beta 37. We note that the behaviors of T yr and Trp residues at the alpha 1-beta 2 interface are not synchronous. It is likely that the behaviors of Tyr residues are controlled by the ligatio n of beta heme through His-beta 92(F8)-->Val-beta 98(FG5)-->Asp-beta 99(G1) -->Tyr-alpha 42(C7) or Tyr-beta 145(HC2).