Biochemical characterization of the Parkinson’s disease-associated deubiquitylase USP30 using its physiological substrates

Gupta Kheskani, Niyati (2024) Biochemical characterization of the Parkinson’s disease-associated deubiquitylase USP30 using its physiological substrates. MSc(R) thesis, University of Glasgow.

Abstract

Mitochondria are essential for eukaryotic life, existing under tightly regulated control mechanisms. Clearance of damaged mitochondria (mitophagy) is a crucial part of mitochondrial homeostasis and relies on the ubiquitination of proteins on damaged mitochondria, which leads to degradation and removal of the damaged organelle. Crucially, dysregulation of mitophagy is among the leading causes of diverse neurodegenerative disorders, including Parkinson’s Disease (PD).

Several deubiquitinating enzymes (DUBs) have gained attention due to their ability to counteract ubiquitination-dependent mitophagy. USP30 is a DUB that emerged as a potential therapeutic target for PD due to its unique position within a mitophagy signalling cascade, whereby USP30 antagonises the heightened mitophagic flux that is common in hereditary forms of PD. Hence, considerable effort has been invested in developing USP30 inhibitors. However, this has been challenging because USP30 substrate recognition is generally poorly understood and there is a dearth of published USP30-inhibitor complex structures available.

By producing a physiologically relevant USP30 substrate, this project aims to develop an in vitro enzyme assay to understand USP30 substrate recognition, as well as examine the inhibition of USP30 by the new sulphonamide derivative inhibitors: MF-094 and Compound 39. This information can also be used to guide future structural analysis of USP30 in complex with one of its physiologically relevant substrates in the presence or absence of available inhibitors for the development of a crystal system from which to develop and design new inhibitors.

• Item Type: Thesis (MSc(R))
• Qualification Level: Masters
• Additional Information: Supported by funding from Ubiquigent.
• Subjects: Q Science > QR Microbiology
• Colleges/Schools: College of Medical Veterinary and Life Sciences > School of Molecular Biosciences
• Supervisor's Name: Walden, Professor Helen
• Date of Award: 2024
• Depositing User: Theses Team
• Unique ID: glathesis:2024-84471
• Copyright: Copyright of this thesis is held by the author.
• Date Deposited: 22 Jul 2024 13:52
• Last Modified: 22 Jul 2024 13:52
• Thesis DOI: 10.5525/gla.thesis.84471
• URI: https://theses.gla.ac.uk/id/eprint/84471