FTIR spectroscopic study of the dynamics of conformational substates in hydrated carbonyl-myoglobin films via temperature dependence of the CO stretching band parameters.
Two hydrated carbonyl myoglobin (MbCO) films, one containing (0.30 g water)/(g MbCO) from MbCO solution in water at pH 5.5 and the other (0.32 g water)/(gMbCO) from 0.1 M potassium phosphate buffer solution at pH 6.8, were studied by FTIR spectroscopy from 293 K to 78 K at selected temperatures on cooling and reheating. Above approximately 180 K the general trend in temperature dependence of half-bandwidths, peak maxima, and band area ratios of the A1 and A3 conformer bands is similar to those reported by Ansari et al. (1987. Biophys. J. 26:337) for MbCO in 75% glycerol/water solution, but abrupt changes in slopes at approximately 180-200 K and freezing-in of conformer populations, which could be taken as indicator for glass transition of the solvent or the protein, are absent for the hydrated MbCO films. This is interpreted in terms of an exceptionally broad distribution of relaxation times, and is in accord with conclusions from recent calorimetric annealing studies of hydrated protein powders (Sartor et al. 1994. Biophys. J. 66:249). Exchange between the three A conformers does not stop at approximately 180-200 K but occurs over the whole temperature region studied. These results are then discussed with respect to MbCO's behavior in the glass-->liquid transition region of glass-forming solvents, and it is concluded that, in analogy to the behavior of low-molecular-weight compounds with a distribution of rapidly interconverting conformers, freezing-in of MbCO's A conformer populations by the solvent should not be mistaken for a glass transition of MbCO.
Mayer
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Institut für Allgemeine, Anorganische und Theoretische Chemie, Leopold-Franzens-Universität Innsbruck, Austria.
D000818:Animals; D006736:Horses; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009132:Muscles / Q000378:metabolism; D009211:Myoglobin / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D017550:Spectroscopy, Fourier Transform Infrared; D013816:Thermodynamics; D014867:Water
D009211:Myoglobin; C018013:carboxymyoglobin; D014867:Water