About the Project
http://www2.mrc-lmb.cam.ac.uk/group-leaders/t-to-z/r-williams
https://www2.mrc-lmb.cam.ac.uk/groups/rlw/
The phosphatidylinositol 3-kinase (PI3K) known as VPS34 is the primordial member of the PI3K family of enzymes that include both lipid and protein kinases. These enzymes function in large protein complexes, and a subset of these enzymes, including class I PI3Ks, VPS34 and mTOR assemble in complexes that are active on cellular membranes. We are using single-particle cryo-electron microscopy, cryo-electron tomography and hydrogen/deuterium exchange mass spectrometry to understand the structures of these complexes, how they assemble on membranes and how they are regulated on membranes by other membrane-associated proteins. These enzyme complexes have key roles in ageing, neurodegeneration, autophagy, phagocytosis, endocytosis and immunity.
The project involves determining how these complexes are recruited to cellular compartments where they are activated by association with G-proteins and/or receptor tyrosine kinases. Assembly of these complexes on membranes is fundamental to their functions and understanding this process could be fundamental for pharmaceutical development. The structural work will be carried out in parallel with mammalian cell biology and in vitro assays. The HDX-MS provides unequalled insight into the interactions that the enzyme complexes make with membranes and G-proteins. This technique has been optimised in the group to provide single-residue resolution and is central to innovative approaches that we have implemented to develop allosteric inhibitors.
References
Rostislavleva, K., Soler, N., Ohashi, Y., Zhang, L., Pardon, E., Burke, JE, Masson, GR, Johnson, C., Jan Steyaert, J., Ktistakis, NT, Williams, RL. (2015) Structure and flexibility of the endosomal Vps34 complex reveals the basis of its function on membranes. Science 350:aac7365. PMID 2645021
Ohashi Y, Soler N, García Ortegón M, Zhang L, Kirsten ML, Perisic O, Masson GR, Burke JE, Jakobi AJ, Apostolakis AA, Johnson CM, Ohashi M, Ktistakis NT, Sachse C, Williams RL (2016). Characterization of Atg38 and NRBF2, a fifth subunit of the autophagic Vps34/PIK3C3 complex. Autophagy 12: 2129-2144. PMID: 27630019
Baretić D, Berndt A, Ohashi Y, Johnson CM, Williams RL (2016). Tor forms a dimer through an N-terminal helical solenoid with a complex topology. Nat Commun. 7:11016. PMID: 27072897
Baretić D. and Williams RL. (2014) PIKKs--the solenoid nest where partners and kinases meet. Curr Opin Struct Biol 29:134-142. PMID 25460276
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