An investigation into the conformational changes and lipid dependence of NTS1 activation by its agonist

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Many of our neurological functions are controlled through receptor proteins (called GPCRs) residing in the brain, and for this reason it has been estimated that about 30% of all drugs we use act on these receptors, of which there are more than 800. The functions controlled by GPCRs are numerous, and one receptor may be involved in many responses. We need to find out how these receptors work, and it has been said that this is the major challenge of current structural biology (Lagerstrom & Schioth, 2008, Nature Reviews Drug Discovery, 7, 339-57). To help in that process of understanding, and as a result of recent breakthroughs, it is now possible to make a very limited number of GPCRs so that we can start to discover how they function. The one we have elected to work on is involved in pain management, neuromuscular diseases such as Parkinson s and the control of appetite, and hence obesity control. We have been able to express this mammalian brain receptor in simple E. coli bacterial cells, and it is functionally active to make any of the work we do relevant to its function in the brain. As with many of these receptors, they are activated by the binding of small molecules, and although we can monitor this binding, the important aspect is to understand and investigate how the protein is then activated and how it sends its signal to other proteins and then ultimately the cell. We will, therefore, express and purify the receptor, activate it with hormone, and then use state-of-the-art methods to probe the receptor changes during the activation process. These methods can be used to measure distances at the nanoscale (0.5-8nm +/- 0.01nm) in these nanodetectors , as well as the time scale (in microseconds - nanoseconds) for the activation process. Since this protein normally sits in a membrane of the brain cell, it needs some of the lipid components of the membrane to function properly, and we will also investigate which of these components are required. Finally, if time permits, we will determine how the activated receptor passes the signal to the next component in the chain of signalling molecules, to unravel a little more about this vital signalling mechanism. All the information from this cutting edge project will add to our general understanding of brain signal transmission, and help in future drug design and disease control.