Developing super-resolution imaging based biophysical methods to study protein aggregate induced membrane disruption at the single-molecule level

This proposal aims to develop a digital, ultrasensitive technique that can detect and quantify protein-induced membrane permeabilisation and disruption at the highest level of sensitivity. Disruption of membrane integrity is a ubiquitous mechanism by which protein aggregates confer toxicity in various human diseases, including Alzheimer's and Parkinson's disease, Type II diabetes, Sickle cell disease, and even specific type of Cancer. Toxic aggregates disrupt and permeabilise lipid bilayer of the cellular membrane and allow the influx of extracellular calcium ions, which leads to disrupted calcium homeostasis and cell death. In this work, we will fabricate nanosized probe to mimic the natural bilayer of cellular membrane and determine how protein aggregate damage membrane integrity. By utilising single-molecule localisation based super-resolution imaging, we will develop a quantitative method which will enable direct observation of individual protein aggregates induced lipid membrane permeabilisation in a high-throughput manner by counting the number of entering ions. This novel ultra-sensitive methodology will provide new insights into the mechanism of membrane permeabilisation in protein-misfolding disease, allowing us to identify and characterise the most toxic protein aggregates in complex human samples and screen the potential drug which can prevent or reduce protein aggregates induced membrane damage which is related to Alzheimer's and Parkinson's disease.