Abstract
The formation of amyloid fibrils is linked to multiple neurodegenerative disorders, including Alzheimer’s and Parkinson’s disease. Despite years of research and countless studies on the topic of such aggregate formation, as well as their resulting structure, the current knowledge is still fairly limited. One of the main aspects prohibiting effective aggregation tracking is the environment’s effect on amyloid-specific dyes, namely thioflavin-T (ThT). Currently, there are only a few studies hinting at ionic strength being one of the factors that modulate the dye’s binding affinity and fluorescence intensity. In this work we explore this effect under a range of ionic strength conditions, using insulin, lysozyme, mouse prion protein, and α-synuclein fibrils. We show that ionic strength is an extremely important factor affecting both the binding affinity, as well as the fluorescence intensity of ThT.
Highlights
Protein aggregation into insoluble, highly structured amyloid fibrils is related to the onset and progression of many neurodegenerative disorders, such as Alzheimer’s or Parkinson’s diseases [1,2]
There are multiple methods used to track protein aggregation into amyloid fibrils. Changes in their secondary structure can be analyzed by circular dichroism [10] or Fourier-transform infrared spectroscopy [11]; aggregate morphology is commonly examined by transmission electron microscopy [12] or atomic force microscopy [13], while changes in fibril quantity are determined by sedimentation or amyloidophilic dye binding [14]
We examine ThT binding to amyloid fibrils formed of either model amyloidogenic proteins–insulin [39] and lysozyme [3], or neurodegenerative disease-related prion protein [40] and α-synuclein [41] under a large range of ionic strength conditions
Summary
Highly structured amyloid fibrils is related to the onset and progression of many neurodegenerative disorders, such as Alzheimer’s or Parkinson’s diseases [1,2]. Very few potential anti-amyloid compounds have passed the initial clinical trials, and none have been approved as effective in treating or curing patients [8,9]. These two factors are intertwined, as a limited comprehension of protein fibrillization and the methods used to track it led to the identification of seemingly potential, yet ineffective, disease-modulating compounds. When examining a potential anti-amyloid compound, these drawbacks could lead to a false interpretation of the results and yield another failed clinical trial
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