Lipid-protein interactions: Characterisation of the oxysterol mediated protein complexes by chemical proteomics

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Cellular function relies on the coordinated communication and action of a collection of biomolecules including proteins and lipids. Lipids exhibit diverse biological effect via their interacting receptors. Proteins that contain a lipid binding domain can, upon binding of their lipid ligand, undergo a conformational change, which leads to interaction with new partners or translocation to another subcellular compartment. Such multicomponent complexes are transient, or dynamic under specific conditions, and difficult to investigate. Conventional biochemical methods to study lipid-protein interactions often require large amounts of material and are carried out on a 'one-at-a-time' basis. Thus, in this proposal we plan to develop a high throughput platform to capture in real time a picture of the spatial and temporal 'global' lipid-protein interaction network using a combination of cross-linking, lipid pull-down, and proteomics technologies. The identified interactions will then be examined for their effect on protein function and cellular activities using a quantitative proteomics approach. The class of lipids will be focused on in this study are oxysterols. Oxysterols are oxidized derivatives of cholesterol and their imbalance is implicated in atherosclerosis and neurodegenerative diseases. We will use the developed novel approach to identify oxysterol receptors and reveal their involvement in protein complexe assembly and disassembly. The results will potentially aid in the elucidation of biological process, the understanding of the importance of oxysterols in the healthy state and how their imbalance may be the cause of disease.