Photophysical Properties of Fluorescent Probe Thioflavin T in Crowded Milieu
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...
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| Format: | Article |
| Language: | English |
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Wiley
2017-01-01
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| Series: | Journal of Spectroscopy |
| Online Access: | http://dx.doi.org/10.1155/2017/2365746 |
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| author | Natalia P. Rodina Maksim I. Sulatsky Anna I. Sulatskaya Irina M. Kuznetsova Vladimir N. Uversky Konstantin K. Turoverov |
| author_facet | Natalia P. Rodina Maksim I. Sulatsky Anna I. Sulatskaya Irina M. Kuznetsova Vladimir N. Uversky Konstantin K. Turoverov |
| author_sort | Natalia P. Rodina |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-da43ce49d2b54804bbe5b8a8dc7aa277 |
| institution | Kabale University |
| issn | 2314-4920 2314-4939 |
| language | English |
| publishDate | 2017-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Spectroscopy |
| spelling | doaj-art-da43ce49d2b54804bbe5b8a8dc7aa2772025-02-03T01:25:54ZengWileyJournal of Spectroscopy2314-49202314-49392017-01-01201710.1155/2017/23657462365746Photophysical Properties of Fluorescent Probe Thioflavin T in Crowded MilieuNatalia P. Rodina0Maksim I. Sulatsky1Anna I. Sulatskaya2Irina M. Kuznetsova3Vladimir N. Uversky4Konstantin K. Turoverov5Laboratory of Structural Dynamics, Stability and Folding of Proteins, Institute of Cytology, Russian Academy of Sciences, Saint Petersburg, RussiaLaboratory of Structural Dynamics, Stability and Folding of Proteins, Institute of Cytology, Russian Academy of Sciences, Saint Petersburg, RussiaLaboratory of Structural Dynamics, Stability and Folding of Proteins, Institute of Cytology, Russian Academy of Sciences, Saint Petersburg, RussiaLaboratory of Structural Dynamics, Stability and Folding of Proteins, Institute of Cytology, Russian Academy of Sciences, Saint Petersburg, RussiaLaboratory of Structural Dynamics, Stability and Folding of Proteins, Institute of Cytology, Russian Academy of Sciences, Saint Petersburg, RussiaLaboratory of Structural Dynamics, Stability and Folding of Proteins, Institute of Cytology, Russian Academy of Sciences, Saint Petersburg, RussiaThioflavin 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.http://dx.doi.org/10.1155/2017/2365746 |
| spellingShingle | Natalia P. Rodina Maksim I. Sulatsky Anna I. Sulatskaya Irina M. Kuznetsova Vladimir N. Uversky Konstantin K. Turoverov Photophysical Properties of Fluorescent Probe Thioflavin T in Crowded Milieu Journal of Spectroscopy |
| title | Photophysical Properties of Fluorescent Probe Thioflavin T in Crowded Milieu |
| title_full | Photophysical Properties of Fluorescent Probe Thioflavin T in Crowded Milieu |
| title_fullStr | Photophysical Properties of Fluorescent Probe Thioflavin T in Crowded Milieu |
| title_full_unstemmed | Photophysical Properties of Fluorescent Probe Thioflavin T in Crowded Milieu |
| title_short | Photophysical Properties of Fluorescent Probe Thioflavin T in Crowded Milieu |
| title_sort | photophysical properties of fluorescent probe thioflavin t in crowded milieu |
| url | http://dx.doi.org/10.1155/2017/2365746 |
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