The Fe(2+)-His(F8) Raman band shape of deoxymyoglobin reveals taxonomic conformational substates of the proximal linkage.
The band shape of the Raman line attributed to the Fe(2+)-N(epsilon)(His(F8)) stretching mode in deoxymyoglobin contains significant information on the nature of the Fe-His proximal linkage. Raman lines appearing close to it, however, obscure the true line profile. To isolate this from its accompanying lines we use its isotopic shift of approximately 1 cm(-1) when (56)Fe in natural-abundance deoxymyoglobin is substituted by (54)Fe. This enables us to isolate the true line shape. We have measured this line shape in sperm whale myoglobin dissolved in a 66% vol/vol glycerol/water solution for nine temperatures from 10 K to 300 K. The nu(Fe-His) band shows a complex temperature-dependent profile, with a shoulder on its high-frequency wing, which becomes more prominent with increasing temperature. Detailed analysis reveals that the band is composed of five distinct lines attributable to taxonomic conformational substates of the nu(Fe-His) linkage. These are in thermodynamic equilibrium above the glass transition temperature T(f) but freeze in into the thermodynamic distribution at T(f) for lower temperatures. Alternative models that try to explain the nu(Fe-His) band shape by either an anharmonic coupling of the nu(Fe-His) to a low-frequency heme doming mode or by conformational substates related to a Gaussian distribution of iron out-of-plane displacements are at variance with the distinct features observed experimentally.
Schott
J
J
FB1-Institut für Experimentelle Physik, Universität Bremen, 28359 Bremen, Germany.
Dreybrodt
W
W
Schweitzer-Stenner
R
R
D000818:Animals; D006639:Histidine / Q000378:metabolism*; D007501:Iron / Q000378:metabolism*; D009211:Myoglobin / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D013059:Spectrum Analysis, Raman; D014907:Whales
D009211:Myoglobin; C055547:deoxymyoglobin; D006639:Histidine; D007501:Iron