Please use this identifier to cite or link to this item:
https://www.um.edu.mt/library/oar/handle/123456789/88432
Title: | Characterisation of xanthine oxidoreductase variants associated with heart disease and hypertension |
Authors: | Cuschieri, Jade (2021) |
Keywords: | Hypertension Heart -- Diseases Uric acid Xanthine oxidase Oxidoreductases |
Issue Date: | 2021 |
Citation: | Cuschieri, J. (2021). Characterisation of xanthine oxidoreductase variants associated with heart disease and hypertension (Bachelor's dissertation). |
Abstract: | Hypertension has been linked with hyperuricaemia and polymorphisms of xanthine oxidoreductase, the protein which catalyses the last two steps of uric acid production. In this study, the A932T variant, a variant of xanthine oxidoreductase which is known to increase the risk of hypertension, was produced as part of a project to determine how the mutation affects the activity, structure and stability of the human xanthine oxidoreductase protein. The wildtype and the A932T variant were expressed in TP1000 cells and purified by immobilised metal affinity chromatography. Native-PAGE and an NBT assay were carried out to determine whether the purified protein samples were active. A uric acid assay was then performed to measure the activity of both the wildtype and the A932T variant and to determine the percentage xanthine oxidase and the percentage xanthine dehydrogenase that were present within the samples. The NBT assay showed that both the wildtype and the A932T variant were active, while the native-PAGE showed that only the wildtype was active. The uric acid assay confirmed that the A932T variant was active but it showed that it was less active than the wildtype. Further optimisation of the expression of the variant needs to be carried out since previous studies have shown that this variant is more active than the wildtype. The purification step also needs to undergo optimisation since the protein samples obtained for both the wildtype and the variant were not pure enough to perform circular dichroism. Therefore, the secondary structure and the stability of the A932T variant were not analysed. |
Description: | B.Sc. (Hons) Med. Biochem.(Melit.) |
URI: | https://www.um.edu.mt/library/oar/handle/123456789/88432 |
Appears in Collections: | Dissertations - FacM&S - 2021 Dissertations - FacM&SPB - 2021 |
Files in This Item:
File | Description | Size | Format | |
---|---|---|---|---|
21BMB007.pdf Restricted Access | 9.67 MB | Adobe PDF | View/Open Request a copy |
Items in OAR@UM are protected by copyright, with all rights reserved, unless otherwise indicated.