Fluorescence-detected stopped-flow kinetic investigation of thioflavin T binding onto the amyloid fibril of lysozyme

doi:10.3969/j.issn.1000-1565.2017.05.006

Abstract

Abstract: Deposition of amyloid fibrils in tissues and organs is relevant to a wide range of devastating human diseases, including Alzheimer’s disease, Parkinson’s disease, and type Ⅱ diabetes. Thioflavin T(ThT)is widely used as a fluorescent probe for qualitative and quantitative detections of amyloid fibrils. Yet the exact kinetic mechanism of ThT binding onto amyloid fibril remains elusive. In this study, we used fluorescence-detected stopped-flow technique to study the binding mechanism of ThT onto the amyloid fibril of hen egg white lysozyme induced by guanidine hydrochloride. We proposed a kinetic binding mechanism to interpret our experimental observations. Our results showed that the binding of ThT onto lysozyme fibril follow a dual-site parallel binding mechanism. We believe our work is beneficial for better understanding of the interaction between fluorescent dye and amyloid fibril.

Key words: thioflavin T, lysozyme, amyloid fibril, kinetics

CLC Number:

O643

MA Gang, QIN Zhe, LIU Siyang, SHANG Yanli, JIA Baohuan. Fluorescence-detected stopped-flow kinetic investigation of thioflavin T binding onto the amyloid fibril of lysozyme[J]. Journal of Hebei University (Natural Science Edition), 2017, 37(5): 476-482.

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