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Molecular Probes: What Is the Range of Their Interaction with the Environment?

H. Lesch ^*, J. Schlichter ^*, J. Friedrich ^* and J. M. Vanderkooi 

^* Physik-Department E14, Lehrstuhl für Physik Weihenstephan, Technische Universität München, 85350 Freising, Germany; and Department of Biochemistry and Biophysics, School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104 USA

Correspondence: Address reprint requests to Josef Friedrich, Technische Universität München, Lehrstuhl für Physik Weihenstephan, Vöttinger Str. 40, 85350 Freising, Germany. Tel.: 49-8161-71-3294; E-mail: j.friedrich{at}lrz.tu-muenchen.de.

We performed pressure-tuning hole-burning experiments on a modified cytochrome c protein in a glycerol/buffer glass. The shift and the broadening of the holes were investigated for various frequencies within the inhomogeneous band. On the basis of a simple model, we were able to estimate the interaction range between chromophore and protein. It is 4.5 Å. The parameters that enter the model are the compressibility, the static mean-square displacement, the inhomogeneous width, and the average spectral shift per pressure. From this result and from our experiments on pressure-induced denaturing, we conclude that water molecules have to be brought very close to the chromophore during the denaturation process.

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