Photophysical Properties of Fluorescent Probe Thioflavin T in Crowded MilieuReport as inadecuate




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Journal of Spectroscopy - Volume 2017 2017, Article ID 2365746, 10 pages - https:-doi.org-10.1155-2017-2365746

Research Article

Laboratory of Structural Dynamics, Stability and Folding of Proteins, Institute of Cytology, Russian Academy of Sciences, Saint Petersburg, Russia

Institute of Physics, Nanotechnology and Telecommunications, Peter the Great Saint-Petersburg Polytechnic University, Saint Petersburg, Russia

Department of Molecular Medicine and Byrd Alzheimer’s Research Institute, Morsani College of Medicine, University of South Florida, Tampa, FL 33612, USA

Correspondence should be addressed to Konstantin K. Turoverov

Received 26 September 2016; Revised 20 November 2016; Accepted 8 December 2016; Published 12 January 2017

Academic Editor: Artem E. Masunov

Copyright © 2017 Natalia P. Rodina et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Thioflavin T ThT is a widely used fluorescent probe of amyloid fibrils, which accompanies many serious neurodegenerative and other diseases. Until recently, examinations of processes of amyloid fibril formation in vitro were conducted in solutions whose properties were significantly different from those found inside the densely packed cells. Such crowded cellular milieu is typically simulated in vitro using concentrated solutions of inert polymers, which do not usually interact with proteins. However, these crowding agents can have a direct effect on the ThT molecule, and this effect must be taken into account. We examined the influence of PEG-400, PEG-12000, and Dextran-70 on the photophysical properties of ThT. It was shown that these crowding agents caused the red shift of the absorption, fluorescence excitation, and fluorescence spectra of ThT. Under these conditions, the increases of the molar extinction coefficient, fluorescence quantum yield, and excitation lifetime of ThT are also observed. However, these changes are significantly less pronounced than those observed for ThT bound to fibrils. It is concluded that, despite some effects of crowding agents on intrinsic fluorescent properties of ThT, this dye can be used as a probe of structure and formation of amyloid fibrils in crowded milieu in vitro.





Author: Natalia P. Rodina, Maksim I. Sulatsky, Anna I. Sulatskaya, Irina M. Kuznetsova, Vladimir N. Uversky, and Konstantin K. Turove

Source: https://www.hindawi.com/



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