Fluorescence Technologies for Monitoring Interactions Between Biological Molecules In Vitro - Progress in Molecular Biology and Translational Science - page 109

CHAPTER FOUR

Fluorescence Technologies for
Monitoring Interactions Between
Biological Molecules In Vitro

Sebastien Deshayes, Gilles Divita
Centre de Recherches de Biochimie Macromol´culaire, Department of Chemical Biology and
Nanotechnology for Therapeutics, CRBM-CNRS, UMR-5237, UM1-UM2, University of Montpellier,
1919 Route de Mende, Montpellier, France

Contents
1. Introduction to Fluorescence 110

1.1 Fluorescence principle 110

1.2 Solvatochromism and resonance energy transfer 111

1.3 Fluorescence for biological molecules 117

2. Monitoring Protein/Substrate Interactions 118

2.1 Protein/nucleotide interactions 118

2.2 Protein/nucleic acid interactions 121

2.3 Peptide/nucleic acids 125

3. Monitoring Protein/Protein and Polypeptide Interactions 128

3.1 Probing protein/protein interactions 128

3.2 Probing peptide/protein interactions 132

4. Fluorescence for Protein/Membrane Interactions 133

4.1 Probing membrane interactions with tryptophan solvatochromism 133

4.2 Probing membrane insertion by tryptophan quenchers 135

4.3 Probing membrane interactions with specific probes 136

5. Conclusions and Perspectives 137

Acknowledgments 138

References 138

Abstract
Over the last two centuries, the discovery and understanding of the principle of
fluorescence have provided new means of characterizing physical/biological/chemical
processes in a noninvasive manner. Fluorescence spectroscopy has become one of
the most powerful and widely applied methods in the life sciences, from fundamental
research to clinical applications. In vitro, fluorescence approaches offer the potential to
sense in real-time extra and intracellular molecular interactions and enzymatic reactions,
which constitutes a major advantage over other approaches to the study of biomolec-
ular interactions. This technology has been used for the characterization of protein/