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Detection of type I collagen fibrils formation and dissociation by a fluorescence method based on thioflavin T.

Authors :
Zou M
Yang H
Wang H
Wang H
Zhang J
Wei B
Zhang H
Xie D
Source :
International journal of biological macromolecules [Int J Biol Macromol] 2016 Nov; Vol. 92, pp. 1175-1182. Date of Electronic Publication: 2016 Aug 09.
Publication Year :
2016

Abstract

In this study, fibrillogenesis and thermal dissociation of pepsin-soluble collagen (PSC), extracted from snakehead (Channa argus) skin, were monitored by fluorescence method based on thioflavin T (Th-T), where the accuracy and sensitivity were evaluated and compared with those of turbidity assay. The fluorescence method revealed the fibrillogenesis dynamics of collagen with better sensitivity, especially at nucleation and plateau stages. The melting temperature (T <subscript>m</subscript> ) of PSC was estimated to be 47°C by circular dichroism spectroscopy; below this temperature, the triple-helical structure should be intact. After that, the dynamic process of collagen dissociation was explored by the fluorescence method, and verified by morphological analysis of the fibrils and the proportion of retained fibrils. The thermal dissociation critical temperature (TDCT) of PSC fibrils was confirmed to be 39°C. The fluorescence intensity of fibril-incorporated Th-T gradually decreases in the dissociation process, and the decrease rate can be accelerated by increasing temperature. Finally, the thermal stability of triple-helical structures of free-, assembled- and dissociated-PSC was compared. Thus, we demonstrated the formation and thermal dissociation of collagen fibrils in vitro by a fluorescence method based on Th-T. This approach may advance the understanding of fibril formation and inverse dissociation of fish-sourced collagen in vitro.<br /> (Copyright © 2016 Elsevier B.V. All rights reserved.)

Details

Language :
English
ISSN :
1879-0003
Volume :
92
Database :
MEDLINE
Journal :
International journal of biological macromolecules
Publication Type :
Academic Journal
Accession number :
27519293
Full Text :
https://doi.org/10.1016/j.ijbiomac.2016.08.027