A comparative pressure tuning hole burning study of protoporphyrin IX in myoglobin and in a glassy host

We measured the behavior of spectral holes under isotropic pressure changes as a function of burn frequency. We compared a protein sample, namely protoporphyrin IX substituted myoglobin in a glycerol/water glass with a sample where the protoporphyrin IX was directly dissolved in a host glass. The differences are remarkable - holes in the pure glass behave as expected for a homogeneous isotropic material. It is the nonlinear frequency dependence of the pressure shift where the deviation of the protein sample is most obvious. These observations signal a correlation between the structures of the dye probe and the structures of the apoprotein. They further show that global parameters of the apoprotein, such as the isothermal compressibility, depend strongly on the associated conformational substates and are subject to unexpected large variations.