Dynamics of the metal-binding domains in the regulation of the Wilson disease protein, ATP7B
| dc.contributor.committeeMember | Stone, Scot | |
| dc.contributor.committeeMember | Wu, Yuliang | |
| dc.contributor.committeeMember | Sanders, David | |
| dc.creator | Patry, Jaala | |
| dc.date.accessioned | 2021-01-15T20:09:10Z | |
| dc.date.available | 2022-01-15T06:05:09Z | |
| dc.date.created | 2020-12 | |
| dc.date.issued | 2021-01-15 | |
| dc.date.submitted | December 2020 | |
| dc.date.updated | 2021-01-15T20:09:11Z | |
| dc.description.abstract | The Wilson disease protein, ATP7B, is a human copper transporter of the P-type ATPase class that is required for maintaining copper homeostasis in the body. ATP7B contains six metal binding domains that possess a ferredoxin-like βαββαβ-fold. Each metal binding domain (MBD) has one copper-binding site characterized by a CxxC motif and each is connected to the next by a flexible linker. MBD1-3 associate through transient interactions, as do MBD5-6. MBD1-3 transitions from a closed conformation to an open conformation upon receiving copper from the copper chaperone ATOX1. Enzymatic activity and intracellular trafficking of ATP7B are governed by the dynamics of the first three cytosolic MBDs. In our study, we used SAXS analysis and model calculation to determine the copper-dependent change in conformational space and MBD arrangement of the MBD1-6 chain. We have also used SAXS and model calculation to determine that residue K269 of MBD3 as essential for proper domain arrangement of the MBD1-3 fragment and its copper-dependent dynamics. We have shown that the compound DC_AC50 (3-amino-N-(2-bromo-4,6-difluorophenyl)-6,7-dihydro-5H-cyclopenta[b]thieno[3,2- e]pyridine-2-carboxamide) is able to bind to MBD1, MBD2, and MBD3. We used molecular docking to identify DC_AC50 as a useful tool to study the copper transport activity and localization of ATP7B. Lastly, we have demonstrated that the molecular basis of Wilson disease caused by the G85V mutation is likely MBD1 misfolding causing ATP7B degradation in the cell. | |
| dc.format.mimetype | application/pdf | |
| dc.identifier.uri | http://hdl.handle.net/10388/13202 | |
| dc.subject | ATP7B | |
| dc.subject | copper transporters | |
| dc.subject | protein structure | |
| dc.title | Dynamics of the metal-binding domains in the regulation of the Wilson disease protein, ATP7B | |
| dc.type | Thesis | |
| dc.type.material | text | |
| local.embargo.terms | 2022-01-15 | |
| thesis.degree.department | Biochemistry | |
| thesis.degree.discipline | Biochemistry | |
| thesis.degree.grantor | University of Saskatchewan | |
| thesis.degree.level | Masters | |
| thesis.degree.name | Master of Science (M.Sc.) |