Centre for structural analysis of complex biological systems

Understanding the function of the molecules of life requires knowledge of their three dimensional structures. Seeing, at or near to the level of individual atoms, how the building blocks of life (proteins and DNA) are assembled enables us to understand both how they may act to drive the chemical reactions that power and maintain living cells, and how they are organised into more complex structures that form the basis of cells and tissues. Detailed knowledge of structure can explain how specific alterations affect function, for example where changes to specific molecules are linked to disease, or how biological systems can be engineered to fulfil useful functions, such as making new drugs or turning carbon dioxide into liquid fuels.

Most structures of biological molecules are derived from experiments where ordered crystals of the pure material are exposed to X-rays. The success of this approach relies upon inducing crystals to form. Unfortunately, for many interesting and important biological molecules this remains very difficult, and large numbers of experiments must be conducted to identify suitable conditions for crystal formation. However, recent technological developments have increased the number of experiments possible with limited amounts of material, and created automated systems to monitor the progress of experiments and detect crystals as they form. Furthermore, technology has improved our ability to create conditions mimicking those existing inside biological membranes (the structures that separate the cell interior from its surroundings and organise the cell into compartments) greatly simplifying the process of obtaining crystals of proteins that are normally associated with membranes. Such proteins perform key biological functions at the cell surface, enabling cells to recognise one another and to bind biological surfaces, and regulating the traffic of molecules, including other proteins, into and out of the cell. However, membrane proteins are much harder to work with, and hence less well understood, than other protein systems.

Here we request funds to purchase equipment that will transform our ability to grow crystals, and obtain structures, of a range of biologically interesting but technically challenging targets. We will create a state-of-the-art Facility to exploit recent successes producing proteins and protein assemblies in the quantities necessary for crystallisation. Specifically, we wish to purchase: i) a robot to set up crystallisation experiments in conditions replicating the membrane environment; ii) an automated system to house the numbers of crystallisation experiments made possible by robotic systems working on small scales, and that will monitor their progress without human intervention; and iii) a complete crystallisation facility, including a robot to set up experiments and a microscope to inspect the results, maintained in a controlled, oxygen-free, environment. We will use this equipment to obtain structures of a number of biological molecules and assemblies including: the machinery controlling protein movement across membranes; the surface proteins of the human red blood cell that determine blood group, surface proteins from disease-causing bacteria that enable them to bind human cells; giant molecular machines synthesising drugs and antibiotics; the protein assembly by which cells carry out the instructions contained within genes; artificial proteins that carry electrons; and a wide range of proteins, involved in processes from bacterial antibiotic resistance to conversion of carbon dioxide into liquid fuels, that only function when oxygen is absent. Through our strong links to other local Universities our Facility, which will be unique within the region, will be open to researchers across the South West and South Wales, and will provide cutting edge instrumentation on which to provide the next generation of scientists with skills essential to the UK science and technology base.

High-resolution structural information can transform our understanding of a given biological system. X-ray diffraction remains the technique with the broadest application to diverse biological targets. Advances in instrumentation at all stages of the macromolecular crystallography pipeline, from protein expression to software tools, have greatly expanded the range of targets that may be considered amenable to structure determination. In this application we seek support to equip a Facility with instrumentation to enable crystallisation of a range of challenging structural targets of high scientific value, including soluble and membrane protein complexes, de novo designed protein systems and oxygen-sensitive enzymes. We request funding for i) a lipidic cubic phase (LCP)-capable robot for crystallisation of membrane proteins and complexes; ii) an automated crystal incubation and imaging system able to maintain and monitor large numbers of experiments, with capability for fluorescence detection of protein crystals and microcrystals in conditions of high background (LCP) and iii) a dedicated anaerobic chamber housing a crystallisation robot and microscope, together with a port for crystal freezing, for crystallisation of oxygen-sensitive systems. Our targets include membrane protein complexes from pro- (protein export; bacterial surface protein) and eukaryotic (erythrocyte membrane proteins) sources, polyketide synthase and eukaryotic transcription super-complexes, de novo designed membrane channel and oxidoreductase systems and a range of radical and redox enzymes. The Facility will enable us to fully exploit recent progress generating material suitable for crystallisation and augment existing capabilities for recombinant protein production and characterisation. No similar equipment is installed in the South West, while our strong local links in structural biology and graduate student training will enable this to operate as a regional Facility with a wide user base.

This proposed Centre will deliver a major new resource for structural biology, advancing our understanding of complex biological systems necessary for medical intervention and their modification and exploitation, which will contribute substantially to the UK's global leadership in these areas. The Centre will progress and diversify such studies, focusing on significant biological challenges that fit with the BBSRC World-Class Bioscience strategy. Moreover the Centre will impact upon important strategic areas of Synthetic Biology and Nanotechnology, by providing new high-resolution information of protein components and complex systems.

We intend that this research will impact upon policy-makers, funding bodies and academic institutions by providing clear evidence of the value of state-of-the-art equipment and fundamental, interdisciplinary research for the future of UK science and education. In order to deliver this impact we will highlight the importance of scientific knowledge in maintaining and increasing the UK's leading position in higher education, emphasising the central role of science in educating the public as well as in schools and lifelong learning. We will also continue to promote such research outside the University, in the South West, nationally and internationally, and discuss potential impact with colleagues, other institutions, funding bodies and learned societies. Moreover by increasing knowledge of basic science the Centre will have a major impact on interdisciplinary training of early-career researchers. Additionally it will facilitate outreach opportunities to educate the public and about the importance of scientific discovery and to inspire young students, revealing the intellectual benefits of a career in bioscience.

The Centre will also foster new projects and collaborations at Bristol and beyond, as well as providing a resource for researchers throughout the UK. It will also provide a hub for workshops and practical courses.

The main areas of impact will be:
a) Delivering a state-of-the-art facility for biological structure determination of complex biological systems, establishing a unique resource in the UK and enabling new collaborations and training opportunities.
b) Delivering scientifically literate people to society. School pupils, undergraduates, postgraduates and early career researchers.
c) Engaging in science festivals, schools visits, talks, debates and press briefings. School pupils have been very effectively involved to date, as have the public; we also plan to formulate new strategies to widen the impact.
d) In the longer term, realising impact in drug development, energy security and synthetic biology.