The core of this project is to contribute to the development of a comprehensive understanding of substrate binding and recognition through the experimental determination of ligand binding sites and poses. Membrane proteins are still a major challenge when considering NMR solution studies, mainly due to their large size and the need to use detergents encumbering the use of protein- observed solution NMR experiments.
In this project a variety of ligand and protein solution NMR experiments will be combined to investigate the transport cycle of LeuT and DAT, which the NeuroTrans consortium can purify in large quantities. Project 2 will be mainly focused on ligand-observed NMR experiments combined with protein-observed experiments using 13C LeuT methyl labelling (A and IVL) in order to establish structure/function relationships and derive information concerning the dynamics of the transport cycle.
The main objectives of project 2 are: