Prof. Anthony Watts

President, European Biophysical Societies Association (EBSA)
Editor, European Biophysical Journal,
Department of Biochemistry
University of Oxford

Title: Multiscale spectroscopies for understanding GPCR mediated cell signalling

Abstract:

G protein-coupled receptors (GPCRs) play a pivotal role in cellular signalling, highlighted by the fact that they form the target for ~40% of marketed pharmaceuticals. While evidence has been accumulating for the existence and functional significance of GPCR oligomers, the matter is still under much debate, in part due to lack of consensus on morphological aspects, such as the receptor interfaces involved in oligomerisation, their possible dynamic nature, and the methods used.

Neurotensin receptor 1 (NTS1) is one of few GPCRs that can be produced in E. coli in an active state, and has been implicated in a variety of conditions including schizophrenia and various cancers. NTS1 has been demonstrated by us to show lipid dependent functionality and dimerise in lipid bilayers, however, there is still no structural or dynamic information on the receptor self-associations in the membrane.

In our recent studies, we combine ensemble FRET, DEER, single molecule methods and in silico approaches to probe NTS1 oligomerization in a membrane. Helix 8, a contentious signalling moiety, has also been investigated in membrane-bound NTS1, using helix scanning using site-directed spin labelling. The bound ligand conformation has been addressed using solid state NMR, and the oligomerization-dependent activation investigated. The results support the presence of a concentration-dependent dynamic equilibrium between monomers and dimers, which could provide a means of regulation of receptor signalling and biased coupling in vivo, and give indication about pathological consequences of receptor associations.

Dijkman et al., (2018) Nature Comms. 9:1710;1-14

 http://www.bioch.ox.ac.uk/~awatts/