Extrinsic fluorescent dyes as tools for protein characterization

Publication Type:

Journal Article

Source:

Pharmaceutical Research, Volume 25, Number 7, pp. 1487-1499 (2008)

ISBN:

0724-8741

DOI Name (links to online publication)

10.1007/s11095-007-9516-9

Keywords:

extrinsic fluorescent dyes; fluorescence spectroscopy; protein aggregation; protein characterization; protein folding; bovine serum-albumin; amyloid fibril formation; intramolecular charge-transfer; molten-globule states; nile-red; thioflavin-t; congo red

Abstract:

Noncovalent, extrinsic fluorescent dyes are applied in various fields of protein analysis, e.g. to characterize folding intermediates, measure surface hydrophobicity, and detect aggregation or fibrillation. The main underlying mechanisms, which explain the fluorescence properties of many extrinsic dyes, are solvent relaxation processes and (twisted) intramolecular charge transfer reactions, which are affected by the environment and by interactions of the dyes with proteins. In recent time, the use of extrinsic fluorescent dyes such as ANS, Bis-ANS, Nile Red, Thioflavin T and others has increased, because of their versatility, sensitivity and suitability for high-throughput screening. The intention of this review is to give an overview of available extrinsic dyes, explain their spectral properties, and show illustrative examples of their various applications in protein characterization.

28/10/2009