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Proceedings Paper

Probing conformational dynamics by photoinduced electron transfer
Author(s): Hannes Neuweiler; Dirk P. Herten; N. Marme; J. P. Knemeyer; Oliver Piestert; Philip Tinnefeld; Marcus Sauer
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Paper Abstract

We demonstrate how photoinduced electron transfer (PET) reactions can be successfully applied to monitor conformational dynamics in individual biopolymers. Single-pair fluorescence resonance energy transfer (FRET) experiments are ideally suited to study conformational dynamics occurring on the nanometer scale, e.g. during protein folding or unfolding. In contrast, conformational dynamics with functional significance, for example occurring in enzymes at work, often appear on much smaller spatial scales of up to several Angströms. Our results demonstrate that selective PET-reactions between fluorophores and amino acids or DNA nucleotides represent a versatile tool to measure small-scale conformational dynamics in biopolymers on a wide range of time scales, extending from nanoseconds to seconds, at the single-molecule level under equilibrium conditions. That is, the monitoring of conformational dynamics of biopolymers with temporal resolutions comparable to those within reach using new techniques of molecular dynamic simulations. We present data about structural changes of single biomolecules like DNA hairpins and peptides by using quenching electron transfer reactions between guanosine or tryptophan residues in close proximity to fluorescent dyes. Furthermore, we demonstrate that the strong distance dependence of charge separation reactions on the sub-nanometer scale can be used to develop conformationally flexible PET-biosensors. These sensors enable the detection of specific target molecules in the sub-picomolar range and allow one to follow their molecular binding dynamics with temporal resolution.

Paper Details

Date Published: 1 July 2004
PDF: 12 pages
Proc. SPIE 5322, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues II, (1 July 2004); doi: 10.1117/12.531167
Show Author Affiliations
Hannes Neuweiler, Ruprecht-Karls-Univ. Heidelberg (Germany)
Dirk P. Herten, Ruprecht-Karls-Univ. Heidelberg (Germany)
N. Marme, Ruprecht-Karls-Univ. Heidelberg (Germany)
J. P. Knemeyer, Ruprecht-Karls-Univ. Heidelberg (Germany)
Oliver Piestert, Ruprecht-Karls-Univ. Heidelberg (Germany)
Philip Tinnefeld, Univ. Bielefeld (Germany)
Marcus Sauer, Ruprecht-Karls-Univ. Heidelberg (Germany)
Univ. Bielefeld (Germany)

Published in SPIE Proceedings Vol. 5322:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues II
Dan V. Nicolau; Joerg Enderlein; Robert C. Leif; Daniel L. Farkas, Editor(s)

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