Developing a nanoplatform to prevent the spread of Parkinson's disease
thesisposted on 2022-03-29, 02:16 authored by India Jade Boyton
Parkinson's disease (PD) spreads in the brain via the release of the protein α-synuclein from diseased neurons into the extracellular space, where nearby healthy brain cells take it up, triggering misfolding and aggregation of α-synuclein in those cells. Thus, pathology spreads in the brain, leading to the symptoms of PD. The overarching aim of this project was to investigate the potential of a novel protein nanocage system for the targeted capture of abnormal α-synuclein, preventing its transmission between brain cells and halting PD progression. Encapsulin protein nanocages selectively self-assemble around proteins tagged with a unique encapsulation signal peptide (ESig), encapsulating them. To better understand this process, the biophysical mechanisms and physicochemical factors underlying encapsulin disassembly/reassembly was characterised using spectroscopic techniques. Encapsulin was found to reversibly disassemble in extreme guanidine hydrochloride (4-7 M) and alkaline conditions and then reassemble within 6-8 hours when returned to normal conditions. Encapsulin disassembly/reassembly conditions were optimised and applied to successfully capture untagged superfolder green fluorescent protein. These results will help pave the way towards the capture of untagged pathological α-synuclein, and the potential future development of a capture system that halts the progression of PD in in vitro models -- abstract.