Volume 8 Issue 1
Feb.  2022
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Long Houfang, Zeng Shuyi, Sun Yunpeng, Liu Cong. Biochemical and biophysical characterization of pathological aggregation of amyloid proteins[J]. Biophysics Reports, 2022, 8(1): 42-54. doi: 10.52601/bpr.2022.210032
Citation: Long Houfang, Zeng Shuyi, Sun Yunpeng, Liu Cong. Biochemical and biophysical characterization of pathological aggregation of amyloid proteins[J]. Biophysics Reports, 2022, 8(1): 42-54. doi: 10.52601/bpr.2022.210032

Biochemical and biophysical characterization of pathological aggregation of amyloid proteins

doi: 10.52601/bpr.2022.210032
Funds:  This work was supported by the Major State Basic Research Development Program (2019YFE0120600), the “Eastern Scholar” project supported by the Shanghai Municipal Education Commission, the Science and Technology Commission of Shanghai Municipality (STCSM) (18JC1420500, 20XD1425000 and 2019SHZDZX02). Figures are created with BioRender.com. We acknowledge the resources provided by this website.
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  • Corresponding author: liulab@sioc.ac.cn (C. Liu)
  • Received Date: 30 July 2021
  • Accepted Date: 17 September 2021
  • Available Online: 17 March 2022
  • Publish Date: 28 February 2022
  • Protein amyloid fibrillation, a process of liquid to solid phase transition, is involved in the pathogenesis of a variety of human diseases. Several amyloid proteins including α-synuclein (α-syn), Tau, amyloid β (Aβ) protein, and TAR DNA-binding protein 43 kDa (TDP-43) form pathological fibrils and deposit in patient brains of different neurodegenerative diseases (NDs) such as Parkinson’s disease (PD), Alzheimer’s disease (AD) and Amyotrophic lateral sclerosis (ALS). Preparation and characterization of amyloid fibrils in vitro are essential for studying the molecular mechanism underlying the dynamic amyloid aggregation and its pathogenesis in diseases. In this protocol, we take PD-associated α-syn as an example, and describe amyloid protein purification and fibrillation approaches. We then introduce biochemical and biophysical characterization of amyloid fibrils by Thioflavin-T (ThT) fluorescence kinetics assay, transmission electron microscopy (TEM), atomic force microscopy (AFM) and multiple fibril stability measurement assays. The approaches described here are applicable to different amyloid proteins, and are of importance for further study on the structure determination of amyloid fibrils and their pathological function in cells and animal models.
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