University of Twente Student Theses
Analyzing the protein complex formation of SARS-CoV-2 nucleocapsid protein and α-Synuclein
Hove, J.H. ten (2024) Analyzing the protein complex formation of SARS-CoV-2 nucleocapsid protein and α-Synuclein.
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| Abstract: | Parkinson’s disease is a neurodenerative diseasse caused by aggregation of α- Synuclein which can be triggered by a viral infection. Research suggest that SARS-CoV-2 could be such a viral infection and initiate amyloid formation causing Parkinson’s disease. To gain insight in the relation between these two diseases on molecular niveau, this thesis aims to investigate the interactions between the nucleocapsis protein of SARS-CoV-2 and α-Synuclein further. The described study attempts to determine the change in protein complex size for the nucleocapsid protein and α-Synuclein using fluorescence correlation spectroscopy. The proteins are labeled and in a low concentration prepared for a fluorescence measurements, where the proteins are detected when they move to the focal volume. This yields a fluorescent intensity trace that can be analyzed with an autocorrelation function to extract the diffusion coefficient of the protein complexes in the sample. The diffusion coefficient of α-Synuclein is found to be 85.22 ± 56.47 μm2/s, where the diffusion coefficient for the nucleocapsid protein is found to be 23.0 ± 2.9 μm2/s. The binding events and affinity between the two proteins for different con- centrations of α-Synuclein are also investigated using microscale thermophoresis. This measures a fixed concentration labeled nucleocapsid protein with different concentrations of an unlabeled ligand protein. This thesis assesses the binding events for both unlabeled nucleocapsid protein and α-Synuclein. The equilib- rium dissociation constant found for the interaction between the labeled and unlabeled nucleocapsid protein is KD = 0.49 − 0.62 μM , where the dissociation constant for the interaction between the SARS-CoV-2 nucleocapsid protein and α-Synuclein turned out to be KD = 3.3122 ± 1.1948 μM . |
| Item Type: | Essay (Bachelor) |
| Faculty: | TNW: Science and Technology |
| Subject: | 02 science and culture in general, 33 physics |
| Programme: | Applied Physics BSc (56962) |
| Link to this item: | https://purl.utwente.nl/essays/97983 |
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