Mitochondria are vital for energy production, cell function and homeostasis, and thereby health. This project explores how the protein PINK1, key to mitochondrial quality-control, is imported into mitochondria and how its misdirection contributes to early-onset Parkinson’s disease. We have discovered a new pathway for PINK1 import, and show that its structure influences its fate and function. Using advanced biochemical, biophysical, and computational tools, the project will investigate how PINK1 variants associated with Parkinson’s affect mitochondrial function, and progression of disease. This interdisciplinary project offers exciting opportunities to uncover novel mechanisms of neuro-degeneration and gain hands-on experience with cutting-edge research techniques.
Background:
Mitochondria are essential organelles responsible for ATP production, metabolic regulation, and biosynthesis. Their function depends on the precise import of nuclear-encoded proteins via the TOM and TIM complexes, particularly the TIM23 pathway. This import is vital for mitochondrial biogenesis and maintenance. A key player in mitochondrial quality control is the kinase PINK1, which under normal conditions is imported, subject to cleavage by the rhomboid protease PARL. This then enables retro-translocation and degradation. However, in damaged mitochondria, PARL cleavage is prevented and PINK1 accumulates on the outer membrane, triggering mitophagy. Mutations in PINK1 disrupt this process and are linked to early-onset Parkinson’s disease.
Recent findings from our laboratories suggest an unexpected alternative import route for PINK1 into the mitochondrial matrix via the TIM23 pathway (see reference below). Structural modelling indicates that PINK1’s trans-membrane domain (TMD) may adopt different conformations, influencing its import fate and function. This discovery opens new avenues for understanding of the regulation of mitochondrial structure, function and quality control, relevant to the progression of Parkinson’s disease.
Key Research Question:
How does the structural flexibility of PINK1’s TMD determine its mitochondrial import pathway and functional role, and how do disease-associated mutations disrupt this process to contribute to the progression Parkinson’s associated neuro-degeneration?
Specific Objectives:
1. Structural and dynamic modelling of PINK1
- Use computational modelling (e.g., AlphaFold, Molecular Dynamics) to predict different conformational states of the TMD, and determine how they can be stabilised.
- Determine how variants of PINK affect the structure of its TMD.
2. Mechanistic Analysis of TIM23 Import and PARL Cleavage
- Establish mitochondrial import assays of PINK1 and examine the effects of variants (identified above) affecting TMD conformation.
- Establish cleavage assays of PINK1 by PARL, and examine the effects of variants (identified above) affecting TMD structure.
3. Consequences of TMD perturbation, and PINK1 fate, on mitochondrial structure and function
- Generate cell lines producing variants of PINK with different TMD structures.
- Assess consequences for mitochondrial function (respiration, membrane potential, reactive oxygen species (ROS)) using Seahorse assays and fluorescence microscopy.
- Examine effects on mitochondrial structure by electron cryo-tomography.
4. Impact of Parkinson’s-linked PINK1 variants
- Introduce disease-associated mutations into PINK1 constructs.
- Analyse effects on PINK1 import and cleavage (as above).
- Monitor the effects on mitochondrial structure and function (as above)
- Correlate molecular defects with disease mechanisms.
Student Ownership and Development:
The student will have the opportunity to:
- Design and optimise experimental protocols. Lead computational modelling and structural validation.
- Select and characterise disease-relevant PINK1 variants.
- Interpret data and propose new hypotheses.
- Contribute to publications and present findings at conferences.
This interdisciplinary project combines computational biology, biochemistry, structural biology and cell biology, offering a rich training environment and the chance to make impactful discoveries in mitochondrial biology and neuro-degeneration.
About the GW4 BioMed2 Doctoral Training Partnership
The partnership brings together the Universities of Bath, Bristol, Cardiff (lead), and Exeter to develop the next generation of biomedical researchers. Students will have access to the combined research strengths, training expertise, and resources of the four research-intensive universities, with opportunities to participate in interdisciplinary and 'team science'. The DTP has already awarded over 90 studentships across 6 cohorts in its first phase, along with 80 students over 4 cohorts in its second phase.
How to apply
Please complete an application to the GW4 BioMed2 MRC DTP for an ‘offer of funding’ on GW4 BioMed MRC DTP
Please complete the online application form linked from the DTP’s website by 5.00pm on Monday, 20th October 2025. Please note that we may close the application process before the stated deadline if an unprecedented number of applications are received– check the DTP’s website for details and updates. If you are shortlisted for interview, you will be notified from Tuesday, 23rd December 2025. Interviews will be held virtually on 27th and 28th January 2026. Studentships will start on 1st October 2026. If successful, you will also need to make an application for an 'offer to study' at University of Bristol. Instructions for doing this will be provided nearer the time.
Application Enquiries
For enquiries relating to the DTP programme or funding, please contact GW4BioMed@cardiff.ac.uk
Please contact the project supervisor for project-related queries.