Biogenesis of mitochondrial complexes
- Bio21 Molecular Science and Biotechnology Institute
|Prof David Stroudemail@example.com|
Summary The Stroud lab uses advanced systems biology techniques to understand the assembly and function of multi-subunit complexes and how their dysfunction underpins a number of important human diseases. We employ a range of mass-spectrometry techniques, including shotgun proteomics, affinity enrichment and BioID proximity studies, metabolic labelling and multi-omics approaches and combine them with patient derived cell lines, CRISPR/Cas9 gene editing, and classical biochemistry. Our research is aimed at both understanding the primary biology underpinning human disease, as well as developing systems biology tools to improve patient diagnostic outcomes.
Is has been estimated that, even at rest, our bodies turn over ~70kg of ATP each day. More than 90% of this is generated through mitochondrial oxidative phosphorylation (OXPHOS), which occurs on the five membrane protein complexes comprising the respiratory chain. Mitochondria are comprised of ~1500 different proteins. Over 80 of these are subunits of respiratory chain complexes and >50 others known as assembly factors are needed for their biogenesis and regulation. Many more proteins support energy production indirectly. Surprisingly, we still don't know the functions of ~200 human mitochondrial proteins! This project is focused on characterising selected mitochondrial proteins of unknown function (see Hock et al., 2020 Mol. Cell. Proteom. as an example). Most of these are implicated in mitochondrial disease, the most common form of inherited metabolic disease, affecting ~1/5000 births.
Here, you will use CRISPR/Cas9 gene-editing to knock out uncharacterised proteins to validate their involvement in mitochondrial energy generation. You will also utilise newly developed affinity enrichment and BioID proximity labelling tools coupled with mass-spectrometry to identify their protein-protein interactions. Shotgun proteomics as well as metabolomics will be used to understand the impact of protein loss on the proteome and metabolome. In this project you will become familiar with mammalian cell culture, molecular cloning, mass-spectrometry and systems biology, Blue-Native PAGE, bioenergetic measurements, and general protein biochemistry techniques.