Probing physical properties of single amyloid fibrils using nanofluidic channels
Journal article, 2023

Amyloid fibril formation is central to the pathology of many diseases, including neurodegenerative disorders such as Alzheimer's and Parkinson's disease. Amyloid fibrils can also have functional and scaffolding roles, for example in bacterial biofilms, and have also been exploited as useful biomaterials. Despite being linear protein homopolymers, amyloid fibrils can exhibit significant structural and morphological polymorphism, making it relevant to study them on the single fibril level. We here introduce the concept of nanofluidic channel analysis to the study of single, fluorescently-labeled amyloid fibrils in solution, monitoring the extension and emission intensity of individual fibrils confined in nanochannels with a depth of 300 nm and a width that gradually increases from 300 to 3000 nm. The change in fibril extension with channel width permitted accurate determination of the persistence length of individual fibrils using Odijk's theory for strongly confined polymers. The technique was applied to amyloid fibrils prepared from the Alzheimer's related peptide amyloid-β(1-42) and the Parkinson's related protein α-synuclein, obtaining mean persistence lengths of 5.9 ± 4.5 μm and 3.0 ± 1.6 μm, respectively. The broad distributions of fibril persistence lengths indicate that amyloid fibril polymorphism can manifest in their physical properties. Interestingly, the α-synuclein fibrils had lower persistence lengths than the amyloid-β(1-42) fibrils, despite being thicker. Furthermore, there was no obvious within-sample correlation between the fluorescence emission intensity per unit length of the labelled fibrils and their persistence lengths, suggesting that stiffness may not be proportional to thickness. We foresee that the nanofluidics methodology established here will be a useful tool to study amyloid fibrils on the single fibril level to gain information on heterogeneity in their physical properties and interactions.

Author

Nima Sasanian

Chalmers, Life Sciences, Chemical Biology

Rajhans Sharma

Chalmers, Life Sciences, Chemical Biology

Quentin Lubart

Chalmers, Life Sciences, Chemical Biology

Sriram Kesarimangalam

Chalmers, Life Sciences, Chemical Biology

Marziyeh Ghaeidamini

Chalmers, Life Sciences, Chemical Biology

Kevin D. Dorfman

University of Minnesota

Elin Esbjörner Winters

Chalmers, Life Sciences, Chemical Biology

Fredrik Westerlund

Chalmers, Life Sciences, Chemical Biology

Nanoscale

2040-3364 (ISSN) 20403372 (eISSN)

Vol. In Press

Vad karaktäriserar en toxisk amyloid oligomer?

Swedish Research Council (VR) (2016-03902), 2017-01-01 -- 2020-12-31.

The Stability of Amyloid Fibrils - a case study on a-synuclein

Swedish Research Council (VR) (2020-05303), 2021-01-01 -- 2024-12-31.

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Swedish Research Council (VR) (2020-03400), 2021-01-01 -- 2024-12-31.

Next Generation Nanofluidic Devices for Single Molecule Analysis of DNA Repair Dynamics

European Commission (EC) (EC/H2020/866238), 2020-04-01 -- 2025-03-31.

Subject Categories

Physical Chemistry

Biochemistry and Molecular Biology

Biophysics

DOI

10.1039/d3nr02740f

PubMed

37953701

More information

Latest update

11/28/2023