A Structure-Function Map of the Interactions of the σ1 Receptor (360G-Wellcome-106275_Z_14_Z)

We aim to generate a protein interaction map for the sigma1 signalling pathway and use this to target biophysical and structural studies. To achieve this aim we will use a concerted in vivo and in vitro approach. Lifetime imaging fluorescence microscopy will be used to monitor interactions by FRETin vivo, with NMR and other structural and biophysical techniques being used to characterise these interactions in vitro. Information from the in vitro analyses will allow for further in cell experiments testing the molecular mechanisms of action of the sigma1 pathway. Initial work with the sigma1– BiP (Binding immunoglobulin Protein) interaction should serve as a test case allowing for the development of methodologies that can be applied to other sigma1 interactions. In the first instance we will expand this approach to the shaker related voltage gated potassium channel family. Starting with Kv 1.4 and Kv 1.5 a fragment based screening approach will be applied to search for points of interaction and look at how sigma1 interacts with and regulates ion channels. This project should not only yield structural information on the sigma1 receptor and its interactions but will also provide a methodological frame work for a systematic dissection of the sigma1 pathway.