Linking Conformation Change to Hemoglobin Activation Via Chain-Selective Time-resolved Resonance Raman Spectroscopy on Protoheme/Mesoheme Hybrids
Journal of Biological Inorganic Chemistry
Time-resolved Resonance Raman spectra are reported for Hb tetramers, in which the αand β chains are selectively substituted with mesoheme. The Soret absorption band shift in meso- relative to protoheme permits chain-selective excitation of heme RR spectra. The evolution of these spectra following HbCO photolysis show that geminate recombination rates and yields are the same for the two chains, consistent with recent results on 15N-heme isotopomer hybrids. The spectra also reveal systematic shifts in the deoxy-heme ν4 and νFe-His) RR bands, which are anti-correlated. These shifts are resolved for the successive intermediates in the protein structure, which have previously been determined from time-resolved UVRR spectra. Both chains show Fe-His bond compression in the immediate photoproduct, which relaxes during the formation of the first intermediate, Rdeoxy (0.07 μs), in which the proximal F-helix is proposed to move away from the heme. Subsequently, the Fe-His bond weakens, more so for the α than the β chains. The weakening is gradual for the β chains, but abrupt for the α chains, coinciding with completion of the R-T quaternary transition, at 20μs. Since the transition from fast- to slow-rebinding Hb also occurs at 20μs, the drop in the α chain νFe-His supports the localization of ligation restraint to tension in the Fe-His bond, at least in the α-chains. The mechanism is more complex in the β chains.
Balakrishnan, Gurusamy; Ibrahim, Mohammed; Mak, Piotr J.; Hata, Jessica; Kincaid, James R.; and Spiro, Thomas G., "Linking Conformation Change to Hemoglobin Activation Via Chain-Selective Time-resolved Resonance Raman Spectroscopy on Protoheme/Mesoheme Hybrids" (2009). Chemistry Faculty Research and Publications. 398.
Accepted version. Journal of Biological Inorganic Chemistry, Vol. 14, No. 5 (June 2009): 741-750. The final publication is available at Springer via http://dx.doi.org/10.1007/s00775-009-0487-7. © 2009 Springer. Used with permission.
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