Structural and electronic factors that influence oxygen affinities: a spectroscopic comparison of ferrous and cobaltous oxymyoglobin.
Various structural and electronic factors that result in similar rates of oxygen association (kon) and differing rates of oxygen dissociation (koff) for ferrous (FeMb) and cobaltous (CoMb) myoglobin have been investigated. Similar values for kon indicate similar barriers to oxygen binding for CoMb and FeMb. Through optical spectroscopy, we have found that the stable quantum yields of photolysis for CoMbO2 (0.55 +/- 0.05) and FeMbO2 (0.50 +/- 0.05) at 10 K are the same. The X-ray absorption near edge spectra (XANES) of CoMb and FeMb reveal similar metal-heme displacements for the deoxy, oxy, and low temperature photoproduct states of CoMb and FeMb. Thus, similar barriers to ligand binding, indicated by similar kon's and photoproduct yields for CoMb and FeMb, correlate with the metal-heme displacements for the oxy, deoxy, and low temperature photoproduct states of CoMb and FeMb. Lower values of koff for FeMbO2 versus CoMbO2 imply different barriers to oxygen release for the two species. X-ray edge positions of CoMb and FeMb indicate a substantial transfer of electron density from the metal to the ligand upon oxygenation. The distribution of electron density throughout the M-O-O moiety differs for CoMbO2 and FeMbO2. Resonance Raman spectroscopy has demonstrated that the Co-O bond is weaker when compared to Fe-O [Tsubaki, M., & Yu, N. T. (1981) Proc. Natl. Acad. Sci., U.S.A. 78, 3581]. We have used photolyzed/unphotolyzed Fourier Transform Infrared (FTIR) difference spectra of CoMb16O2, CoMb18O2, FeMb16O2, and FeMb18O2 to show that the dioxygen stretching frequency, v(O-O), in CoMbO2 (approximately 1138 cm-1) is higher than FeMbO2 (approximately 1131 cm-1). The dioxygen stretching frequency in CoMbO2 is closer to that of heme protein models lacking a hydrogen bond to the distal histidine, suggesting that formation of the hydrogen bond in FeMbO2 provides a greater effect on the distribution of electron density throughout the Fe-O-O... HN moiety, potentially stabilizing a more ionic Fe-O-O bond. These findings demonstrate important electrostatic differences in the distal environments of CoMbO2 and FeMbO2, resulting in different barriers to oxygen release.
Miller
L M
LM
Department of Physiology and Biophysics, Albert Einstein College of Medicine, Yeshiva University, Bronx, New York 10461, USA.
Chance
M R
MR
D000818:Animals; D003035:Cobalt / Q000737:chemistry / Q000378:metabolism*; D003080:Cold Temperature; D004583:Electrons; D005296:Ferrous Compounds / Q000737:chemistry / Q000378:metabolism*; D006736:Horses; D008786:Metmyoglobin / Q000737:chemistry / Q000378:metabolism*; D009211:Myoglobin / Q000737:chemistry / Q000378:metabolism*; D010100:Oxygen / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D013057:Spectrum Analysis / Q000379:methods
D005296:Ferrous Compounds; D009211:Myoglobin; C028048:ferrylmyoglobin; C017595:oxymyoglobin; D008786:Metmyoglobin; D003035:Cobalt; D010100:Oxygen