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We have studied the effect of high hydrostatic pressure and temperature onthe steady state fluorescence anisotropy of Green Fluorescent Protein (GFP). Wefind that the fluorescence anisotropy of GFP at a constant temperaturedecreases with increasing pressure. At atmospheric pressure, anisotropydecreases with increasing temperature but exhibits a maximum with temperaturefor pressure larger than 20 MPa. The temperature corresponding to the maximumof anisotropy increases with increasing pressure. By taking into account of therotational correlation time changes of GFP with the pressure-temperaturedependent viscosity of the solvent, we argue that viscosity increase withpressure is not a major contributing factor to the decrease in anisotropy withpressure. Our results suggest that the decrease of fluorescence anisotropy withpressure may result from changes in H-bonding environment around thechromophore.

Pressure and temperature dependence of fluorescence anisotropy of Green Fluorescent Protein