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NMR structures of membrane proteins, complexes and lipid assemblies; towards a dedicated wide-bore ultra high field mas spectrometer for biological research


The objective is to prepare a demonstration project aiming at the improvement and implementation of novel ultra high field Magic Angle Spinning NMR technology required for comprehensive structure determination using eg. multidimensional correlation spectroscopy. The demonstration project aims at the construction of a fully operational prototype spectrometer with the neccessary dedicated low-temperature high-speed MAS probes, operating at ultra high field (around 750 MHz) with excellent long-term stability, and the construction of the associated wide-bore ultra high field wide-bore magnet, in a unique collaboration between scientific key laboratories in Europe and the only major European manufacturer of NMR spectrometers. The projected instrument would be used to determine structures in space and time of biomolecules with the desired functions in a multidisciplinary approach of site-specific or multiple isotope labeling of genuine subcellular entities like membrane proteins, complexes and lipid assemblies as defined in area 6.1 structural biology, research tasks 6.1.1 6.1.2 (prominent) and 6.1.3. The long-term goal is to develop flexible technological means to such a level that comprehensive molecular information at atomic resolution can be obtained for widely different types of molecules or subjects, eg. an essentially unperturbed membrane protein receptor when in its natural environment in the membrane, and relate this to function. The preparation stage explores the technical feasibility of the design of the dedicated probes and wide-bore magnet.
Multidimensional homonuclear and heteronuclear test-experiments using multiple 13C labeled and 2H diluted samples will help to determine the prototype design targets with respect to long term instrument stability, temperature range, rf power requirements, magnet specifications and, finally, optimization of existing rf technology for ultra high field MAS.
The final deliverable of the preparatory stage will be a detailed plan on how to combine the most advanced knowledge from the industrial and scientific key-players in Europe into a multidisciplinary enterprise to design unique and widely applicable MAS NMR technology for biological studies, including a strategy of how to make the new instrumentation accessible to all biological solid-state NMR users in Europe, for instance in the form of facilities that would be open for all, and taking into account crucial requirements in the field of isotope chemistry; molecular biology methods for labeling and reconstitution; and NMR development. Economic benefits will accrue for health related industries from using the equipment for gaining urgent knowledge of ligand (eg: drug)-receptor binding site. With access to such information rationale design of biomolecules with the desired functions (eg. drugs) becomes a reality (task 6.1.3). Biotechnological benefits will accrue from forging the pathway for the determination of integral membrane protein structure. NMR manufacturing industry will benefit from a new product brought into production for a potentially developing market.

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Rijksuniversiteit Leiden
2300 RA Leiden

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