Professor Corinne Spickett
Supervisor Details
Scientific Inspiration
Prof Mike Murphy, MRC Mitochondrial Biology Unit – he does fantastic fundamental science on redox stress in mitochondria that has broad biomedical implications while being amazingly unassuming (as well as funny).
Research Groups
Project Details
Prof Spickett is the primary supervisor on the below project:
Understanding the redox regulation of the tumour suppressor and cell signalling protein PTEN
Secondary Supervisor(s): Dr Alex Cheong (Pharmacy)
University of Registration: Aston University
BBSRC Research Themes:
- Understanding the Rules of Life (Immunology, Structural Biology, Systems Biology)
- Integrated Understanding of Health (Ageing)
Apply here!
Deadline: 23 May, 2024
Project Outline
Redox balance is a central aspect of cellular function that regulates cell behaviour across many organisms from microbes to mammals. Cells maintain a balance between oxidizing species (such as hydrogen peroxide or superoxide) and antioxidants; changing the balance in either direction affects macromolecular interactions, gene expression, cell differentiation, proliferation or death. Inflammation and mitochondrial dysfunction can lead to oxidative stress through the release of reactive oxygen species. These responses are important in ageing and metabolic dysfunctions, as well as diseases. Many proteins are “redox-sensitive”: i.e. their functions and activity are affected by oxidative modification of the protein.
Phosphatase and tensin homologue (PTEN) is a tumour suppressor and dual specificity phopsphatase. Through its phospholipid phosphatase activity it antagonizes the PI3K/Akt pathway, altering metabolic balance, limiting cell proliferation, and preventing the blocking of apoptosis. As a protein phosphatase, it dephosphorylates proteins involved in cell motility and migration, also counteracting oncogenic changes. PTEN is known to be redox sensitive, and my group demonstrated that oxidation and modification by lipid oxidation products alters both its activity and its protein interactome. We have also been investigating redox-induced changes in subcellular localization, which are likely to control cell behaviour. However, questions remain over the types of modifications that can cause these changes, the mechanisms involved, and their downstream effects on cell behaviour.
The overall aim of the project is to understand the effects of redox-dependent post-translational modifications on PTEN cell signalling processes that underlie ageing, inflammation. Building on our previous research, we will use a broad, in vitro interactomics approach to determine the effect of oxidative and nitrosative stresses on PTEN and analyse its protein-protein interactions. The interactions will be validated in cellulo along with functional tests to confirm their effects on cell differentiation and proliferation. This work will be carried out in cells expressing native PTEN and PTEN containing mutations in the active site and resolving cysteines, to test the effect of blocking the redox response. The redox-dependent networks will be modelled for different cellular conditions with pathway mapping. In parallel, high resolution microscopy will be used to monitor the effects on subcellular localization and correlate with the interactome and cell function data. This will generate novel information on how the dual activities of PTEN function together in physiological and pathophysiological situations, as well as building a paradigm for integrated redox regulation at the whole-cell level.
References
- Approaches to Investigating the Protein Interactome of PTEN. Smith SL, Pitt AR, Spickett CM. J Proteome Res. 2021; 20(1):60-77.
- Short-chain lipid peroxidation products form covalent adducts with pyruvate kinase and inhibit its activity in vitro and in breast cancer cells. Sousa BC, Ahmed T, Dann WL, Ashman J, Guy A, Durand T, Pitt AR, Spickett CM. Free Radic Biol Med. 2019 144:223-233.
- Reversible oxidation of phosphatase and tensin homolog (PTEN) alters its interactions with signaling and regulatory proteins. Verrastro I, Tveen-Jensen K, Woscholski R, Spickett CM, Pitt AR. Free Radic Biol Med. (2016) Jan 90: 24-34
Techniques
- Mammalian cell culture and bacterial culture.
- Biochemical techniques: cell culture, enzymatic assays, protein modification, enrichment and characterization, and molecular biology (mutagenesis and generation of fusion proteins).
- Analytical techniques: liquid chromatography and mass spectrometry (LC-MSMS), PAGE and western blotting.
- Computational techniques: quantitative data analysis, protein identification (proteomics, Mascot), lipid and oxidized lipid identification (Progenesis and other software), pathway mapping
Prof Spickett is also a co-supervisor on a project with Dr Rebecca Jones.
Previous Projects
Prof Spickett was the primary supervisor on the below project last year:
Rusty cells: redox regulation of cell signalling and responses to stress
Co-supervisor: Dr Alex Cheong (Pharmacy) or Dr Alfred Fernandez-Castane (Engineering)
University of Registration: University of Aston
Project outline:
Redox balance is a central aspect of cellular function that regulates cell behaviour across many organisms from microbes to mammals. Cells maintain a balance between oxidizing species (such as hydrogen peroxide or superoxide) and antioxidants; changing the balance in either direction affects macromolecular interactions, gene expression, cell differentiation, proliferation or death. These responses are important in a wide variety of fields including antimicrobial resistance, responses to environmental toxins, cell manufacturing (BRIC) and effects of ageing, diet and health. While the general principles are well understood, many questions remain about the precise interactions that control specific processes. My group works on understanding the sensing and effects of redox imbalance at a molecular level in a variety of systems and applications.
The role of redox balance in magnetotactic bacteria (Co-supervisor Dr Alfred Fernandez-Castane). Magnetotactic bacteria produce magnetosomes, structures that contain magnetite (Fe3O4) and allow the microbe to align in magnetic fields. Magnetosomes are of great interest as sources of magnetic nanoparticles for biotechnology applications, but knowledge of their synthesis, regulation and biochemical effects is still limited. Evidence is emerging that their production is redox-dependent, they may be redox-active in vivo and have protective effects against environmental stress. Building on expertise in growth of magnetotactic bacteria at Aston, this project will investigate the properties of the magnetosome membrane under different growth and redox conditions to determine how the bacteria avoid lethal oxidative damage from iron-dependent reactions and what antioxidant activities are present. Lipidomic and proteomic studies under different conditions will provide new understanding of magnetosome properties and enable improved magnetosome production.
References:
Development of a simple intensified fermentation strategy for growth of Magnetospirillum gryphiswaldense MSR-1: physiological responses to changing environmental conditions. Alfred Fernández-Castané, Hong Li, Owen RT Thomas, Tim W Overton. New Biotechnol 2018; 46: 22-30
Analysis of SMALP co-extracted phospholipids shows distinct membrane environments for three classes of bacterial membrane protein. Teo ACK, Lee SC, Pollock NL, Stroud Z, Hall S, Thakker A, Pitt AR, Dafforn TR, Spickett CM, Roper DI.
Sci Rep. 2019; 9(1):1813.
Lipid Composition Analysis Reveals Mechanisms of Ethanol Tolerance in the Model Yeast Saccharomyces cerevisiae. M Lairón-Peris, S J Routledge, J A Linney, J Alonso-Del-Real, C M Spickett, A R Pitt, J M Guillamón, E Barrio, A D Goddard, A Querol. Appl Environ Microbiol. 2021; 87(12):e0044021.
BBSRC Strategic Research Priority: Renewable Resources and Clean Growth - Industrial Biotechnology, Understanding Rules of Life - Microbiology, Structural Biology, and Systems Biology, and Integrated Understanding of Health - Ageing, and Diet and Health.
Techniques that will be undertaken during the project:
This project will use a combination of the biochemical, analytical and computational techniques below.
- Biochemical techniques: cell culture, enzymatic assays, protein modification, enrichment and characterization, and molecular biology (mutagenesis and generation of fusion proteins).
- Analytical techniques: liquid chromatography and mass spectrometry (LC-MSMS), PAGE and western blotting.
- Computational techniques: quantitative data analysis, protein identification (proteomics), lipid and oxidized lipid identification (Progenesis and other software), pathway mapping.