Assembly of the Commander complex
- 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.
Sorting of proteins and lipids into and out of the endomembrane system is crucial for maintaining cellular homeostasis. Defects in the machineries that control this process lead to a number of debilitating diseases, including neurodegeneration, hypercholesterolemia and Wilson’s disease. Recently, a new and evolutionarily conserved receptor recycling complex known as Commander was discovered. We and others propose that Commander is an endosomal sorting machinery required for the transport and homeostasis of vital plasma membrane receptors.
The principles that govern the assembly of the 16 Commander subunits into the multisubunit complex however, as well as the overall mechanism by which this complex regulates endosomal trafficking in health and disease remains completely unknown. In this project you will use CRISPR/Cas9 to knock out Commander subunits and accessory factors in human cell lines. You will perform shotgun quantitative proteomics and Blue-Native PAGE to understand the effect of subunit loss on complex assembly. Affinity enrichment and BioID proximity labeling coupled to mass-spectrometry will be used to probe protein-protein interactions. In addition, you will become proficient in molecular cloning, confocal microscopy, protein biochemistry and human cell culture techniques.