Interactions between DNA and the Parkinson’s disease protein, -synuclein
Ladda ner
Typ
Examensarbete för masterexamen
Master Thesis
Master Thesis
Program
Biotechnology (MPBIO), MSc
Publicerad
2018
Författare
Westling, Alvina
Modellbyggare
Tidskriftstitel
ISSN
Volymtitel
Utgivare
Sammanfattning
With an increasing life expectancy, a deeper knowledge of neurodegenerative age-related diseases, such as Parkinson’s disease (PD), has become of great importance. The main pathological feature of PD is the degeneration of neuromelanin-containing neurons (NCN) in the part of the brain controlling the involuntary movement of skeletal muscles. The degeneration of the NCN are shown to be linked to toxic aggregation species of -synuclein ( S), which is a protein found in the brain and is believed to regulate the synaptic activity in the brain. S has also been observed in the cytoplasm and nucleus and there are suggestions that structural changes can be induced in S by binding with DNA, but there are only a few studies on S-DNA interactions. This thesis therefore aims to obtain a deeper understanding of the role of protein-DNA interactions in vitro, to enable for the possibility that S-DNA interactions may have some participation in PD. For example, in relation to the gene regulation. To investigate the interactions, synthesised wild-type S has been studied in bulk phase as well as at a single molecule level, with four types of commercially available DNA sequences. In studies in the bulk phase, with lin-ear dichroism, interactions between protein and DNA were observed. Secondary structural changes in S and DNA was not evident in linear dichroism and circu-lar dichroism experiments. To investigate the protein-DNA interactions at a single molecule level, nanofluidic channels and atomic force microscopy were used. It was then observed that the interactions between S and DNA induce a sti˙er configu-ration in the DNA molecules, causing an elongation in length of 30%. The increase in length of the DNA molecules after interactions with S could be of interest when further investigating the gene-regulating role of S. Images of the interactions also revealed that the binding of S to DNA appears in small clusters along the DNA molecule, suggesting that there are favourable S binding sites on the DNA molecule.
Beskrivning
Ämne/nyckelord
Biokemi och molekylärbiologi , Biofysik , Medicinsk bioteknologi (med inriktning mot cellbiologi) , Livsvetenskaper , Biochemistry and Molecular Biology , Biophysics , Medical Biotechnology (with a focus on Cell Biology) , Life Science