Shedding Light on the Photoactivation of Visual Pigments - A Study Using FTIR Spectroscopy

F. de Lange

Promotor: J.J.H.H.M. de Pont
Copromotor: W.J. de Grip
Radboud University Nijmegen
October 11, 1999

Abstract

The work presented in this thesis demonstrates that FTIR difference spectroscopy, in combinationwith protein engineering, stable-isotope labeling, and ligandmodification, is a very sensitive tool to probe the conformational rearrangements leading to receptor activation in visual pigments. Not only does it facilitate the identification of structurally active groups within the complex, it also allows the nature of the structural changes and intramolecular interactions to be studied. It has further been demonstrated that polarized infrared difference spectroscopy in the ATR-mode yields additional resolving power and that it can be fruitfully applied to obtain mechanistic information on the relative orientation of functionally important groupswithin the ligand-protein complex. Clearly, the applicability of this approach is not limited to the case of visual pigments. With the achievement of large-scale functional expression of other (G-protein coupled) receptors and the availability of photo-activatable caged ligands, (ATR-)FTIR is expected to provide invaluable information on the mechanism of activation in a variety of other receptor systems as well.

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