Expression and characterization of human proteins involved in neurological disorders

My Ph.D. research project, entitled “Expression and characterization of human proteins involved in neurodegenerative diseases”, was focused on the application of molecular biology and proteomics methodologies to prepare samples of proteins involved in neurodegenerative diseases. The target proteins were α-synuclein (α-syn) and the amyloid-beta peptides (Aβ), involved in the pathogenesis of Parkinson’s disease (PD) and Alzheimer’s disease (AD), respectively. The aim of my research activity was the optimization of the expression and purification of the neurodegeneration-associated proteins to carry out the following projects:  “NMR analysis of the aggregation kinetics of Aβ1-40”, to reveal aggregation mechanisms of Aβ1-40 and to develop a kinetic model describing the formation of oligomeric and fibrillary species.  “NMR analysis of the assembly of Aβ42: Aβ40 mixed fibrils”. This project is in the frame of an integrative study aimed to characterize the structure of the mixed fibrils (containing Aβ1-42 and Aβ1-40 peptides) at atomic detail.  “Development of a protein aggregation assays for the diagnosis of synucleinopathies” This project was focused on the development of a protocol tool for the diagnosis PD, dementia with Lewy bodies (DLB) and multiple system atrophy (MSA) based on α-synuclein aggregation assays (SAA-seeding aggregation assays) starting from aliquots of Cerebrospinal fluid (CSF).  Study of the interaction between alpha-synuclein and human biofluids components” This project concerned the study of the interaction of α-synuclein with lipoproteins, proteins and other constituents of CSF and plasma.