EAST-NMR

EAST-NMR (Enhancing Access and Services to East European users Towards an efficient and coordinated Pan-European pool of NMR capacities to enable global collaborative research & boost technological advancements) is a European Union research project funded by the Seventh Framework Programme (FP7) that started February 1st 2009 and will run for 48 months. The project type is Combination of a Collaborative Project & Coordination and Support Action (CP&CSA, former I3). The total budget of EAST-NMR is € 3.6 million of which € 3.5 million is funded by EU FP7. This equals total funded effort of 33 person years. The funds are divided between three activities. 19% goes to Networking, 27% to Transnational Access and 38% to Joint Research Activities. Other related EU funded projects are EU-NMR and eNMR.

Motivation

Nuclear Magnetic Resonance (NMR) spectroscopy is a pre-eminent technique for the provision of information on the three-dimensional structures of biomolecules. NMR has continuously advanced its capability for solving the structures of molecules of increasing complexity (macromolecules), notably proteins and nucleic acids, and complexes thereof over the last two decades. NMR experiments reveal the structural and dynamic properties of macromolecules (and their adducts) in solution or additionally, in the last years, in the solid-state. Solid-state NMR spectroscopy has successfully been applied to determine structures of bound ligands binding to their native receptors in the native environment and breakthroughs have been obtained for [ion channels]. European research teams directly collaborate with scientific groups with primary expertise in molecular and cell biology that provide samples for experimental measurements carried out by the NMR spectroscopists. The outcomes of these studies are of interest for an even wider community of scientists, as a result of the biological and biomedical implications of the obtained results. These advancements in structural biology provide key insights into the reactions inside our cells, and are of paramount importance in translational research towards new [...] development.

Usage of NMR in Europe

The European bio-NMR community is composed of many hundred research groups ranging in size from a few to several tens of individual researchers tackling a broad portfolio of different biological problems. The entire community of stakeholders therefore includes several thousand scientists across Europe. The running I3 project EU-NMR is focused on the provision of NMR measurement time and scientific expertise to European researchers, who need access to the best available equipment and most sophisticated know-how for their experiments. Approximately one-hundred projects are carried out yearly through this access scheme at the EU-NMR research infrastructures (RIs). Since its existence in FP3, thousands of researchers have made use of the European biomolecular NMR improvements. In addition to the EU-NMR project, the EC-funded Coordination Action NMR-Life brings together leading bio-NMR research groups throughout Europe. This partnership is committed to closely monitoring the development of bio-NMR techniques and technologies, with a particular focus on aspects relevant for the investigation of macromolecular complexes.

Current access to NMR Research Infrastructures in Europe

Statistics from the on-going EU-NMR project demonstrate that a large imbalance exists in the requests for NMR machine time originating from Eastern Europe and Western Europe. Western Europe has traditionally benefited from a global leadership role in NMR spectroscopy, while, despite the presence of excellent scientists, NMR has failed to reach its full potential as a tool for the study of biomolecules in the East. Western European scientists are increasingly specializing in tackling more challenging projects, in part because of the initial use of the many mid-sized instruments available at universities throughout Western Europe. A growing fraction of the users applying to the EU-NMR project have already screened their samples locally for feasibility and routine measurements; the large RIs provide the highest level of service and access for the most sophisticated spectrometer and experimental schemes requested. Unfortunately, a historical lack of NMR in the East has led to a diminished interest in the tool itself due to a lack of understanding of its potential applications and capabilities, perpetuating a vicious cycle in which this powerful analytical tool is being drastically underused.

Access to solid-state NMR in Europe

The rapid developments in solid-state NMR have been noticed by a considerable number of scientists in the field. However, the number of groups that are capable of providing access and expertise at the frontiers of this rapidly developing field is limited. Information resulting from surveys of the NMR and broader Molecular Life Science communities and input from (potential) users at the EU-NMR user meetings has signalled the urgency of providing access to solid-state NMR. Since the start of the EU-NMR initiative, solid-state technology has significantly evolved, and new instruments have been added to those previously available at the EU-NMR RIs. Specifically, a 900 MHz narrow bore magnet has been added in CNRS-CRNM, and 850 MHz wide bore magnets at CERM and BMRZ, there equipped with a cryogenically cooled probe. These instruments reinforce the capabilities provided by the EU-NMR RIs and provide new opportunities for the field for all European scientists.

NMR protein preparation

Modern NMR spectroscopy relies on the investigation of proteins, RNA, and DNA that are enriched with NMR-active, stable isotopes (13C, 15N, 2H). Such samples are produced by biochemical methods. Currently, the heterologous expression of proteins in Escherichia coli and the transcription of RNA/DNA can be considered as state-of-the-art technologies. [...]-edge investigations rely on the availability of such molecules, which constitutes the major bottleneck for NMR investigations in the Life Sciences. However, many proteins cannot be expressed in bacterial cells but must be prepared in eukaryotic cell lines, or by cell-free protein synthesis. This situation is typically encountered for the preparation of membrane proteins. The unavailability of standardised expression protocols renders NMR spectroscopic investigations difficult for many biology groups.

Solution

The EAST-NMR project responds to the new challenges facing NMR and its Research Infrastructures. Based on the I3 model EAST-NMR gathers major academic and industrial stakeholders to provide solutions in the form of 1) Networking, 2) Transnational Access and 3) Joint Research Activities. The combination of the three elements in an appropriate manner will ensure the necessary advancements for NMR RIs at a pan-European level in terms of enhancing their services to researchers and encouraging the conduction of top quality research. The project will provide transnational access to NMR instrumentation based in Eastern Europe and solid-state NMR facilities, educate and train researchers in NMR’s potential and use advanced sample preparation technologies especially of difficult to tackle membrane proteins through joint research activities.

Networking Activities

The networking activities are focused on good practices (exchange of protocols and best practices through a website, communication with experts, twinning of young scientists, lab rotation), towards the users (focused national/regional conferences, structured training on the theory and practice of NMR, workshops on sample preparation) and towards virtual research communities (web site, connection to Grid e-infrastructure). Young investigators meetings and promoted scientific talks (within the conferences) will be organised.

Transnational Access Activities

A set of eight NMR research infrastructures will be offering transnational access to European research groups under the EAST-NMR Integrated Activity:

The access offered by the Eastern RIs (SLONMR, NCBR, DEBNMR, WARNMR) is expected to be of high impact both for these four RIs and for scientific groups based in Eastern Europe working in the field of bio-NMR. The four RIs will act as nuclei for spreading awareness about the potential of NMR spectroscopy for solving challenging programs among research groups in their regions, in providing users with first-hand experience in accessing such instrumentation. They will actively interact with the RIs BMRZ, SONNMRLSF, CERM, which will specifically provide access to solid-state NMR spectrometer (to analyse e.g. membrane proteins), and UOXF (that will specialize on the characterisation of membrane associating proteins by liquid-state NMR).

The RIs in EAST-NMR will provide the following to visiting scientists:

• Use of advanced methods in NMR spectroscopy for determining the 3D structure of biomolecules and complexes of these molecules in solid and solution states
• Special solid-state NMR hardware to carry out experiments involving the entire range of NMR active nuclei
• High-spinning speed probe equipment for MAS NMR
• Implementation and testing of methods for generating the structure of biomolecules on the basis of experimental and semi-empirical data
• Access to software for data processing, structure determination, and other specific purposes (analysis of kinetics of protein ligand interactions, relaxation analysis, relaxation dispersion analysis, specialised tools for metabolomics, processing tools for fast recording techniques)
• Support by customising software for particular needs of users
• The use of extensive support facilities for sample generation, characterisation and purification as well as for computer modelling
• Support by qualified, dedicated staff throughout all stages of the project and related experiments
• Guidance and support, if required, in the preparation of the research proposal and its experimental details for obtaining access to the RI
• Guidance and support, if required, in transforming the experimental results into a paper
• On request, active participation of the staff of the RI in the research project
• Advanced training opportunities for all aspects of structural biology in the various graduate research training programs

Joint Research Activity

The objective of the Joint Research Activity (JRA) is to develop procedures for the production of membrane protein samples for NMR spectroscopy. The JRA aspires to comprehensive, uniform documentation to reach this goal. The offering of optimised procedures for the production of membrane proteins assures an efficient use of the participating NMR research infrastructures from the very early stages of sample preparation and contributes to Good Laboratory Practices (GLP). In addition, structure determination of molecular complexes and increasingly large molecules requires the establishment of robust technologies that provide novel long-range structure restraints including paramagnetic tagging and novel alignment media. This means improvements in both the service offered by the RIs to their users in terms of quality and quantity, and also in the joint use of the RIs.

Partners

The consortium of EAST-NMR comprises 21 academic and industrial partners working together to achieve a pan-European pool of NMR capacities. Eight NMR Research Infrastructures from Slovenia, Czech Republic, Germany, United Kingdom, Poland, Netherlands, Italy and Hungary (Partners 1-8) make the transnational access possible, especially to solid-state NMR, for a large group of research groups in Eastern Europe. In addition, the consortium comprises nine partners (12-20) highly active in networking activities focused on the East and two industrial partners (10-11) acting in the Joint research Activity together with the academic partners involved in the research (1, 6, 7 & 9). Partner 21 provides support to the project coordinator and the consortium in the management, administration and dissemination issues of the project.

1. Johann Wolfgang Goethe-Universität Frankfurt a.M., Center for Biomolecular Magnetic Resonance (Frankfurt a.M., Germany)
2. National Institute of Chemistry, Slovenian NMR Centre (Ljubljana, Slovenia)
3. Masaryk University Brno, National Centre for Biomolecular Research (Brno, Czech Republic)
4. University of Debrecen, Institut of Chemistry (Debrecen, Hungary)
5. University of Warsaw (Warsaw, Poland)
6. Universiteit Utrecht (Utrecht, The Netherlands)
7. Consorzio Interuniversitario Risonanze Magnetiche di Metalloproteine Paramagnetiche, Magnetic Resonance Center (Florence,Italy)
8. The Chancellor, Masters and Scholars of the University of Oxford, Department of Biochemistry (Oxford, United Kingdom)
9. Centre National de la Recherche Scientifique, Centre Européen de RMN à Très Hauts Champs de Lyon (Lyon,France)
10. Asla Biotech Ltd. (Riga, Latvia)
11. Bruker BioSpin GmbH (Rheinstetten, Germany)
12. University of Patras, Department of Pharmacy (Patras, Greece)
13. Eötvös Loránd University, Laboratory of Structural Chemistry and Biology (Budapest,Hungary)
14. Institute of Biochemistry and Biophysics, Polish Academy of Sciences (Warsaw, Poland)
15. Weizmann Institute of Sciences, Department of Structural Biology (Rehovot, Israel)
16. Tallinn University of Technology (Tallinn, Estonia)
17. Latvian Institute of Organic Synthesis, Department of Physical Organic Chemistry (Riga, Latvia)
18. Institute of Chemistry, Slovak Academy of Sciences (Bratislava, Slovakia)
19. Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences (Sofia, Bulgaria)
20. TÜBİTAK Marmara Research Center Food Institute (Gebze Kocaeli, Turkey)
21. eurelations AG (Zurich, Switzerland)

See also

• European Union
• Seventh Framework Programme
• Nuclear Magnetic Resonance

External links

• EAST-NMR
• EU-NMR
• eNMR