USA - Solution structure of large proteins and protein-ligand complexes by high-field NMR

The acquisition of high-resolution conformational, dynamical and ligand binding data in solution requires the application of state-of-the-art NMR, which complements information from X-ray crystallography. This structural information is necessary for the u nderstanding of the function and regulation of the protein systems and their complexes and to attain rational target-directed drug design. We have recently become interested in the complex formation by the ubiquitous regulatory and signalling proteins cal led 14-3-3 with various phosphorylated partners. This is relevant to understand the mechanism of 14-3-3 action in health and disease, with particular reference to their involvement in serotonin and catecholamine function by stabilization of the limiting e nzymes for the synthesis of these neurotransmitters, i.e. tyrosine hydroxylase (TH) and tryptophan hydroxylse (TPH), as well as the role of 14-3-3 proteins in tumor progression. We have thus initiated a multidisciplinary project that includes several youn g members of our group in collaboration with other groups at UiB to undertake the structural characterization of normal and mutant forms of the seven human isoforms of 14-3-3 complexed with several proteins including TH and TPH. This is expected to reveal new diagnostic and therapeutic approaches. Relevant achievement for this project has been the successful tipple (15N, 13C and 2H) labeling of 14-3-3g and our research group has taken out preliminary NMR spectra in our 600 MHz-NMR equipment in Bergen. We have in addition approached Professor Angela Gronenborn (Chair of Department of Structural Biology, University of Pittsburgh, USA and NMR expert) to establish a formal collaboration and to access the high-field NMR equipment located at her lab. This is es sential for the success of the project since high-field NMR (> 750 MHz), which is not available in Bergen (or Norway), is required for the study of the large 14-3-3 proteins (> 250 amino acids) and their complexes.