Doherty, CPA orcid.org/0000-0001-5685-4716, Young, LM, Karamanos, TK et al. (4 more authors) (2018) A peptide‐display protein scaffold to facilitate single molecule force studies of aggregation‐prone peptides. Protein Science, 27 (7). pp. 1205-1217. ISSN 0961-8368
Abstract
Protein aggregation is linked with the onset of several neurodegenerative disorders, including Parkinson's disease (PD), which is associated with the aggregation of α‐synuclein (αSyn). The structural mechanistic details of protein aggregation, including the nature of the earliest protein–protein interactions, remain elusive. In this study, we have used single molecule force spectroscopy (SMFS) to probe the first dimerization events of the central aggregation‐prone region of αSyn (residues 71–82) that may initiate aggregation. This region has been shown to be necessary for the aggregation of full length αSyn and is capable of forming amyloid fibrils in isolation. We demonstrate that the interaction of αSyn71‐82 peptides can be studied using SMFS when inserted into a loop of protein L, a mechanically strong and soluble scaffold protein that acts as a display system for SMFS studies. The corresponding fragment of the homolog protein γ‐synuclein (γSyn), which has a lower aggregation propensity, has also been studied here. The results from SMFS, together with native mass spectrometry and aggregation assays, demonstrate that the dimerization propensity of γSyn71‐82 is lower than that of αSyn71‐82, but that a mixed αSyn71‐82: γSyn71‐82 dimer forms with a similar propensity to the αSyn71‐82 homodimer, slowing amyloid formation. This work demonstrates the utility of a novel display method for SMFS studies of aggregation‐prone peptides, which would otherwise be difficult to study.
Metadata
Item Type: | Article |
---|---|
Authors/Creators: |
|
Copyright, Publisher and Additional Information: | (c) 2018 The Authors Protein Science published by Wiley Periodicals, Inc. on behalf of The Protein Society. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/) |
Keywords: | Single molecule force spectroscopy, alpha-synuclein, gamma-synuclein, aggregation, amyloid, mass spectrometry |
Dates: |
|
Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Structural Molecular Biology 2 (Leeds) |
Funding Information: | Funder Grant number EU - European Union 322408 |
Depositing User: | Symplectic Publications |
Date Deposited: | 09 Feb 2018 10:57 |
Last Modified: | 08 Feb 2019 01:38 |
Status: | Published |
Publisher: | Wiley-Blackwell |
Identification Number: | 10.1002/pro.3386 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:127258 |