Authors: Neuweiler H.; Sauer M.

Source: Current Pharmaceutical Biotechnology, Volume 5, Number 3, June 2004 , pp. 285-298(14)

Publisher: Bentham Science Publishers

Abstract:

This mini-review describes how single-molecule sensitive fluorescence resonance energy transfer (FRET) and photoinduced electron transfer (PET) reactions can be successfully applied to monitor conformational dynamics in biopolymers. Single-pair 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. That is, the monitoring of conformational dynamics of biopolymers with temporal resolutions comparable to those within reach using new techniques of molecular dynamic simulations. 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.

Keywords: conformational dynamics; protein folding; single-molecule spectroscopy; photoinduced electron transfer; dna hairpins; pet-biosensors

Document Type: Review article

DOI: http://dx.doi.org/10.2174/1389201043376896

Affiliations: 1: Fakultat fur Physik, Angewandte Laserphysik und Laserspektroskopie, Universitat Bielefeld, Universitatsstr. 25, 33615 Bielefeld, Germany.

Publication date: 2004-06-01

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• Current Pharmaceutical Biotechnology aims to cover all the latest and outstanding developments in Pharmaceutical Biotechnology. Each issue of the journal contains a series of timely in-depth reviews written by leaders in the field covering a range of current topics in both pre-clinical and clinical areas of Pharmaceutical Biotechnology. Current Pharmaceutical Biotechnology is an essential journal for academic, clinical, government and pharmaceutical scientists who wish to be kept informed and up-to-date with the latest and most important developments.

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