Optical Structural Analysis of Individual α‐Synuclein Oligomers |
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| Authors: | Dr Juan A Varela Dr Margarida Rodrigues Dr Suman De Patrick Flagmeier Dr Sonia Gandhi Prof Christopher M Dobson Prof David Klenerman Dr Steven F Lee |
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| Institution: | 1. Department of Chemistry, University of Cambridge, Cambridge, UK;2. Department of Molecular Neuroscience, Institute of Neurology, University College London, London, UK;3. UK Dementia Research Institute, University of Cambridge, Cambridge, UK |
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| Abstract: | Small aggregates of misfolded proteins play a key role in neurodegenerative disorders. Such species have proved difficult to study due to the lack of suitable methods capable of resolving these heterogeneous aggregates, which are smaller than the optical diffraction limit. We demonstrate here an all‐optical fluorescence microscopy method to characterise the structure of individual protein aggregates based on the fluorescence anisotropy of dyes such as thioflavin‐T, and show that this technology is capable of studying oligomers in human biofluids such as cerebrospinal fluid. We first investigated in vitro the structural changes in individual oligomers formed during the aggregation of recombinant α‐synuclein. By studying the diffraction‐limited aggregates we directly evaluated their structural conversion and correlated this with the potential of aggregates to disrupt lipid bilayers. We finally characterised the structural features of aggregates present in cerebrospinal fluid of Parkinson's disease patients and age‐matched healthy controls. |
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| Keywords: | amyloid fibrils fluorescence anisotropy neurodegeneration Parkinson's disease protein aggregation |
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