Objective Membrane protein transporters catalyze fundamental physiological processes in all kingdoms of life. A schematic view of membrane transport is emerging from a growing number of crystal structures. However, several fundamental features of membrane protein transport are still evasive, such as how the protein transition between states, what triggers these structural rearrangements, and how the release of energy is coupled to the transport event. The proposed research will build an interdisciplinary platform, which uses molecular biology, biophysical and computational techniques, to characterize membrane protein transport over the full spatio-temporal scale. Molecular dynamics (MD) simulations will capture high-resolution dynamics on the nano-to-microsecond timescale, which will identify communicative salt-bridge/H-bond networks, putative transport pathways, and the energetics involved in substrate transport. Predictions from the simulations will be verified by in vitro characterization of mutagenic phenotypes. Time-resolved wide-angle X-ray scattering (TR-WAXS) will monitor the micro-to-millisecond, large-scale structural rearrangements, which are out of reach for the MD simulations. The proposed platform for characterization of membrane protein transporters will be developed using a copper-transporting P-type ATPase (CopA) as the initial model system. The P-type ATPase family is divided into types I-V, depending on the transported substrate. CopA is the first crystal structure from the type-I ATPases, making it a particularly exciting model system. The applicants’ preliminary molecular dynamics (MD) simulations and the CopA transmembrane architecture strongly indicate a unique mode of transport in type-I ATPases. Hence, by means of visualization of copper transport, this proposal enables the development of an interdisciplinary European platform for multifaceted characterization of membrane transport that will be extendable to a range of membrane protein targets. Fields of science engineering and technologymaterials engineeringcrystalsnatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsnatural sciencesbiological sciencesmolecular biology Programme(s) FP7-PEOPLE - Specific programme "People" implementing the Seventh Framework Programme of the European Community for research, technological development and demonstration activities (2007 to 2013) Topic(s) FP7-PEOPLE-2013-CIG - Marie-Curie Action: "Career Integration Grants" Call for proposal FP7-PEOPLE-2013-CIG See other projects for this call Funding Scheme MC-CIG - Support for training and career development of researcher (CIG) Coordinator KUNGLIGA TEKNISKA HOEGSKOLAN EU contribution € 100 000,00 Address BRINELLVAGEN 8 100 44 Stockholm Sweden See on map Region Östra Sverige Stockholm Stockholms län Activity type Higher or Secondary Education Establishments Administrative Contact Monica Thorén (Mrs.) Links Contact the organisation Opens in new window Website Opens in new window Total cost No data