Biophysical approaches to characterising protein-protein interactions and intermediate species in protein aggregation
type: CreativeWork
description: The phenomenon of protein misfolding and aggregation has been associated with over 50 human diseases, including Parkinson’s disease (PD). While the hallmark deposits in aggregation-associated diseases are primarily composed of fibrillar species, intermediate oligomeric species that form during the aggregation process are believed to be a major cause of toxicity. Such species are relatively poorly characterised due to several challenges that render them inaccessible to most conventional techniques; oligomers are only present at extremely low concentrations in the aggregation reaction, and are additionally highly heterogeneous and often transient in nature. In this thesis, I present complementary approaches to address these difficulties. Firstly, an ensemble of stable, kinetically trapped oligomers with varying biophysical characteristics was established, enabling detailed structure-toxicity relationships to be investigated. Secondly, a method for the simultaneous single-molecule level characterisation and fractionation of oligomeric species under native conditions was established and applied to studying intermediate species in Parkinson’s disease-associated protein aggregation. Finally, I present a general approach to optimising experimental design, which maximises both the efficiency and information gain of experiments, and demonstrate its validation and application to several experimental systems.
abstract: The phenomenon of protein misfolding and aggregation has been associated with over 50 human diseases, including Parkinson’s disease (PD). While the hallmark deposits in aggregation-associated diseases are primarily composed of fibrillar species, intermediate oligomeric species that form during the aggregation process are believed to be a major cause of toxicity. Such species are relatively poorly characterised due to several challenges that render them inaccessible to most conventional techniques; oligomers are only present at extremely low concentrations in the aggregation reaction, and are additionally highly heterogeneous and often transient in nature. In this thesis, I present complementary approaches to address these difficulties. Firstly, an ensemble of stable, kinetically trapped oligomers with varying biophysical characteristics was established, enabling detailed structure-toxicity relationships to be investigated. Secondly, a method for the simultaneous single-molecule level characterisation and fractionation of oligomeric species under native conditions was established and applied to studying intermediate species in Parkinson’s disease-associated protein aggregation. Finally, I present a general approach to optimising experimental design, which maximises both the efficiency and information gain of experiments, and demonstrate its validation and application to several experimental systems.
author: Xu, Catherine
language: eng
keywords: Protein aggregation, Amyloid, Oligomers, Alpha-synuclein, Microfluidics
license: https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/14d3dc54-efd7-4541-be62-5d4d0dd8a420/download
license: https://www.rioxx.net/licenses/all-rights-reserved/
date published: 2020-09-29
date modified: 2026-07-22T22:24:17Z
provider: Cambridge University
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: University of Cambridge

PropertyValue
property ID: DOI
value: 10.17863/CAM.65349