Renewal of the 600 MHz solid-state NMR console for biological applications

Lead Research Organisation: University of Warwick
Department Name: Chemistry

Abstract

Solid-state NMR is uniquely positioned to provide atomic resolution structural and dynamic information on complex biological systems that is highly complementary to other structural biology and biophysical techniques. It is used by researchers in the life and physical sciences to address complex problems in systems ranging from proteins to cells and even intact plants.
This in turn leads to advancements in key areas from sustainable and resilient agriculture, safe nutritious food, new pharmaceuticals and better health to new low-carbon 'greener' energy, materials and everyday products.
In this proposal we request to replace an obsolete and often malfunctioning solid-state NMR console with a new state-of-the-art console and two accompanying NMR probes in order to be able to perform a wide range of experiments not only faster but also on more challenging and complex systems and thus tackle a wider set of challenges related to the important societal issues. NMR probes exist in a variety of different specifications, notably with different rotor diameters as well as the number and tuning range of the rf channels - allowing different isotopes to be studied, e.g., hydrogen, carbon, nitrogen and fluorine - and different probes are best suited to different applications. The added value of the upgrade is that there is a number of other NMR probes acquired over time that could be used with the new console and benefit from presented by it improvements. This equipment will be used in a multiuser and multidisciplinary environment where scientists from different disciplines work together in order to maximise a chance of solving important problems that cannot be solved with a single "magic bullet" scientific approach.
Another important aspect of this proposal is that it will enable us to train a new generation of scientists on a state-of-the-art equipment. PhD and postdocs, who would benefit from such training, could carry the skills and the knowledge they acquired as a result of it as independent scientists in academic or industrial setting and apply it to tackling new challenges.
Finally, we also propose to make available to the wider scientific community part of the time on this specialised instrument for "taster" sessions during which they can test whether this methodology can provide answers to their scientific problems. Because of the highly specialised nature of solid-state NMR the end users would be supported by and could tap into the high-level expertise of the local scientists. If longer projects would be required, they could be pursued as collaborations with the experienced local scientists.

Technical Summary

We propose to renew the > 12 years old obsolete 600 MHz SSNMR rf console with a new state-of-the-art console together with the acquisition of new 3.2 mm HXY E-free and 1.3 mm HFX magic angle spinning probes, to enable molecular level characterisation of structure and dynamics of a wide range of biological systems from proteins to cells and intact plants in a multiuser collaborative environment. The new console will result in improvements in sensitivity (especially for 1H detected experiments often required for applications to complex systems), stability (through implementation of the 2H lock), increase of information content (e.g. measuring several experiments at the same time using multiple receivers) and enabling a range of modern experiments not possible or not practical with the current console. The 3.2 mm E-free probe will enable structural and dynamical measurements on hydrated and salty samples such as intact plants. The 1.3 mm HFX probe will enable a wide range of experiments for characterising structure and dynamics in large protein complexes by leveraging the general lack of fluorine background in biological systems and long-range nature for distance restraints involving fluorine. The added value of the upgrade is that it will also bring new life to the existing unique probes available at the 600 MHz instrument leveraging previous investments from EPSRC and BBSRC and provide access to them for the wider community. These include the only in the UK 0.8 mm MAS Samoson probe spinning up to 100 kHz with superior sensitivity due to the largest sample volume from the available 100 kHz MAS designs and a double rotation (DOR) probe providing unique capabilities for quadrupolar nuclei, e.g. 17O.
We propose a mechanism where up to 5% of the instrument time would be dedicated to provide access to the wider scientific community to this specialised instrumentation and solid-state NMR expertise at University of Warwick in a form of short solid-state NMR "tasters".

Planned Impact

The proposed renewal of 600 MHz solid-state NMR spectrometer will deliver impact for the UK academic and industrial sectors and the wider society by enabling new interdisciplinary science in the BBSRC remit that is well-aligned with its strategic priorities but also, in part, supporting state-of-the-art research in a wider context, especially EPSRC remit. In the recognition of the latter fact EPSRC has declared 30% support for this bid.
New technologies and tools impact on key areas from sustainable and resilient agriculture, safe nutritious food, new pharmaceuticals and better health to new low-carbon 'greener' energy, materials and everyday products. To understand living systems and how they function, can often require access to methodologies that provide atomic scale information, such as NMR. From numerous research projects enabled by the proposed equipment specific examples include studies of intact plants and studies of biological systems involved in producing new bioactive compounds. The insight into the molecular architecture of plant cell walls from solid-state NMR provides new knowledge for the development of improved strategies for use of plant materials for biorefining and for breakdown of plant biomass for second generation bioenergy. It also supports the strategic priority of food security by providing knowledge for understanding animal feed digestibility and reducing food waste. The second specific example of research enabled by the proposed instrumentation is motivated by a desire to better understand the biosynthesis of natural products with important applications in medicine and agriculture, and to devise improved strategies for engineering the biosynthetic pathways for such molecules to produce novel derivatives (an important sub-discipline of synthetic biology). The ultimate goal is to create new products with biological activities ranging from antibacterial, antifungal, anticancer, cholesterol-lowering, or immunosuppressant, to herbicidal or insecticidal.
In addition, the instrumentation will help to generate insight for systems and issues of direct technological importance to as reflected, e.g. in support for this bid from industrial project partners, AstraZeneca and GlaxoSmithKline. The spectrometer will support other active collaboration with industrial partners including Johnson Matthey, Infineum, Pfizer, NovoBiotic and Syngenta. Further engagements with SMEs involved in drug discovery activities will be pursued in collaboration with Medicines Discovery Catapult.
Another impact resulting from this project is training a generation of PhD students and PDRAs in state-of-the-art solid-state NMR, who can disseminate and apply their knowledge through independent academic careers, and/or contribute to developing the UK's portfolio of new and existing pharmaceutical, agrochemical and biotechnology companies.
 
Description (PANACEA) - A Pan-European Solid-State NMR Infrastructure for Chemistry-Enabling Access
Amount € 4,998,891 (EUR)
Funding ID 101008500 
Organisation European Commission 
Sector Public
Country European Union (EU)
Start 08/2021 
End 08/2025
 
Description Solid state and solution NMR spectroscopy and cryo-electron microscopy methodology for the characterisation of aggregation mechanisms in proteins
Amount £102,026 (GBP)
Funding ID BB/V50967X/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 08/2020 
End 09/2024
 
Description Access to fast spinning methodology for SMEs in drug discovery 
Organisation Medicines Discovery Catapult
Country United Kingdom 
Sector Private 
PI Contribution We provide access and expertise in fast magic angle spinning solid-state NMR to SMEs in the context of drug discovery projects.
Collaborator Contribution Purchased a 0.7mm magic angle spinning NMR probe to be used for the projects. Provide project management.
Impact Pilot project on characterisation of a docking domain in non-ribosomal peptide synthesase involved in biosynthesis of antibiotic tyrocidine. https://md.catapult.org.uk/case-studies/protein-interactions-in-non-ribosomal-peptide-synthetases-nrpss/
Start Year 2017
 
Description Characterisation of secondary structure of peptides and proteins 
Organisation AstraZeneca
Country United Kingdom 
Sector Private 
PI Contribution Development of NMR and FTIR methodology.
Collaborator Contribution Expertise on solution NMR and FTIR.
Impact No outcomes yet.
Start Year 2021
 
Description Collaboration with pharmaceutical industry (600 MHz BBSRC--funded console, from 2021) 
Organisation AstraZeneca
Country United Kingdom 
Sector Private 
PI Contribution Solid-state NMR analysis of pharmaceutical compounds relevant to companies (AstraZeneca, GlaxoSmithKline, Pfizer) supported by PhD funding and an Innovate UK award (with AstraZeneca)
Collaborator Contribution co-supervision of PhD projects, research ideas
Impact publications are in preparation
Start Year 2021
 
Description Collaboration with pharmaceutical industry (600 MHz BBSRC--funded console, from 2021) 
Organisation GlaxoSmithKline (GSK)
Department GlaxoSmithKline, Stevenage
Country United Kingdom 
Sector Private 
PI Contribution Solid-state NMR analysis of pharmaceutical compounds relevant to companies (AstraZeneca, GlaxoSmithKline, Pfizer) supported by PhD funding and an Innovate UK award (with AstraZeneca)
Collaborator Contribution co-supervision of PhD projects, research ideas
Impact publications are in preparation
Start Year 2021
 
Description Collaboration with pharmaceutical industry (600 MHz BBSRC--funded console, from 2021) 
Organisation Pfizer Ltd
Country United Kingdom 
Sector Private 
PI Contribution Solid-state NMR analysis of pharmaceutical compounds relevant to companies (AstraZeneca, GlaxoSmithKline, Pfizer) supported by PhD funding and an Innovate UK award (with AstraZeneca)
Collaborator Contribution co-supervision of PhD projects, research ideas
Impact publications are in preparation
Start Year 2021
 
Description Development of fast magic angle spinning instrumentation 
Organisation Tallinn University of Technology
Department Technomedicum
Country Estonia 
Sector Academic/University 
PI Contribution Evaluated and provided feedback about various generations of 0.8 mm MAS probe for application on biomolecules.
Collaborator Contribution Provide experimental 0.8mm MAS probe.
Impact (1) Lamley, J. M.; Iuga, D.; Öster, C.; Sass, H.-J.; Rogowski, M.; Oss, A.; Past, J.; Reinhold, A.; Grzesiek, S.; Samoson, A.; Lewandowski, J. R. J. Am. Chem. Soc. 2014, 136 (48), 16800.
Start Year 2012
 
Description Dynamics of b1AR receptor 
Organisation University of Basel
Department Biozentrum Basel
Country Switzerland 
Sector Academic/University 
PI Contribution We performed pilot study of feasibility of probing dynamics of b1AR receptor by solid-state NMR.
Collaborator Contribution Provided isotopically labelled samples.
Impact na
Start Year 2019
 
Description Ice dynamics in the presence of antifreeze molecules 
Organisation University of Warwick
Country United Kingdom 
Sector Academic/University 
PI Contribution Performed SSNMR experiments and analysis
Collaborator Contribution Provide samples and complementary experiments
Impact Elucidated molecular basis for antifreeze properties of PVA (publication under preparation).
Start Year 2018
 
Description Liquid-liquid phase separation in gene expression 
Organisation University of Cambridge
Department Department of Biochemistry
Country United Kingdom 
Sector Academic/University 
PI Contribution We perfrom SSNMR experiments.
Collaborator Contribution The collaborators supply samples.
Impact Currently we only have preliminary experiments suggesting feasibility of the approach.
Start Year 2021
 
Description NMR studies of plants 
Organisation University of Cambridge
Department Department of Biochemistry
Country United Kingdom 
Sector Academic/University 
PI Contribution Run NMR experiments and analysed data.
Collaborator Contribution Provided samples.
Impact Multidisciplinary collaboration.
Start Year 2019
 
Description NMR studies of wood 
Organisation Scion
Country New Zealand 
Sector Public 
PI Contribution Run NMR experiments and analysed data.
Collaborator Contribution Provided 13C-enriched samples.
Impact na
Start Year 2020
 
Description Protein aggregation 
Organisation GlaxoSmithKline (GSK)
Country Global 
Sector Private 
PI Contribution We are developing NMR based methods for characterising protein aggregation.
Collaborator Contribution Exchange of expertise and help with identifying research targets.
Impact Currently only preliminary results obtained.
Start Year 2020
 
Description Structures of antibiotic-lipid II complexes 
Organisation University of Warwick
Country United Kingdom 
Sector Academic/University 
PI Contribution We have used combination of solution and solid-state NMR (including 100 kHz spinning methodology) to solve structures of antibiotics in complexes with lipid II to inform rational drug development efforts.
Collaborator Contribution Synthesize lipid II.
Impact No outcomes yet. Publication in preparation.
Start Year 2015