Core Support for Collaborative Research in Glycobiology

The genome sequencing projects of the past two decades have yielded the startling revelation that the total number of genes in humans is not very different from many model organisms such as worms and plants. This discovery served to highlight the fact that substantial amplification of genomic information occurs after genes are translated into proteins. Glycosylation, which involves the addition of sugars to selected protein amino acid side-chains, is the most abundant and arguably the most important of these post-translational modifications. Indeed, all cells are coated with a sugar-rich layer called the glycocalyx. Chains of sugars, called glycans, on the periphery of the glycocalyx bind to specific sugar-recognition proteins, called lectins, on partner cells. Many important biological processes depend on the ability of cells to appropriately communicate with each other via these sugar-lectin interactions and to respond accordingly. For example, lectins on the surfaces of viruses and bacteria are known to recognise sugars on target cells and attach to them as the first step of infection. Conversely the adaptive immune system is triggered when sugars on the surfaces of pathogens bind to lectins expressed by cells of the host immune system. Although these examples of glycan-lectin recognition are now relatively well understood, many biological processes that are likely to be similarly controlled by glycan-lectin interactions remain enigmatic. For example, how do immune cells in the gut distinguish between beneficial and harmful microbes, and how does a developing foetus escape rejection by the mother despite being half "foreign"? Pivotal to learning how glycans function in these processes is knowing their structures. We specialize in glycan structural determination using a sophisticated analytical technique called mass spectrometry. We do this in collaboration with dozens of scientists from many biological disciplines both nationally and internationally, who are working with us to explore glycan-lectin structure/function relationships. We are particularly interested in characterizing the glycans that are involved in host microbe interactions, in immune regulation and in mediating recognition events during mammalian reproduction. As well as contributing to fundamental understanding of glycan function, we aim to provide structural data to underpin the identification of targets for new drugs and vaccines for the control of pathogens and parasites. We anticipate that our studies of the glycobiology of mammalian fertilization and reproduction will open up new avenues for natural contraception as well as helping infertile couples. Another of our goals is to understand the pathways involved in regulating glycosylation. This is important, not only because altered glycosylation is associated with many chronic health and aging problems, but also because this knowledge will assist the efficient production of well defined glycoprotein biopharmaceuticals. As well as being important constituents of glycoproteins, sugars are the major building blocks of the polysaccharides that constitute the walls of plant cells. There is an urgent need to better understand the biological processes involved in plant cell wall assembly if the potential of biomass as a renewable energy resource and an alternative to fossil fuels for the production of high value chemicals is to be fully realised. We aim to work with plant scientists in the UK and Australia to elucidate the molecular mechanisms of cell wall polysaccharide biosynthesis. Our analytical methods generate large volumes of complex data, so another of our objectives is to create glycoinformatic tools to assist and speed up data interpretation and annotation, and these tools will be made available to the scientific community. We will also develop and populate openly available databases with our glycan structural data, a resource that will benefit scientists from many disciplines around the world.

We request core support for our world-class mass spectrometry laboratory to enable us to continue to undertake glycobiological research of relevance to the BBSRC priority areas of basic bioscience underpinning health, food security and bioenergy. We are working with more than thirty collaborators in the UK and abroad with the overarching goal of providing a molecular understanding of (i) biological events in which glycan-lectin recognition plays a central role, and (ii) regulation of glycosylation pathways in the biosynthesis of eukaryotic and prokaryotic glycoproteins, bacterial lipopolysaccharides and plant cell wall polysaccharides. A key objective of our research is the continued enhancement of our glycomic and glycoproteomic platforms which are based on the integration of ultra-high sensitivity mass spectrometry with carbohydrate and protein chemistry. We aim to continue to develop open source informatic tools for improving data analysis and to substantially improve and populate public data resources. We will apply our technologies to projects in the following areas: (a) the glycobiology of host/pathogen interactions including characterising glycoconjugates of pathogens and parasites with an important aim being to identify potential vaccine candidates; (b) defining the roles of glycans in regulating host-virus interactions, including characterisation of viral glycoproteins as well as the glycan and lectin receptors of their hosts; (c) exploring how host glycosylation influences the gut microbiome thereby contributing to research into human nutrition and health; (d) investigating the regulation of mammalian glycosylation via the study of metabolic dysfunction; (e) contributing to the glycobiology of mammalian fertilization and reproduction including hunting for the egg binding protein on human sperm and defining structure/function relationships in glycodelin glycoforms; (f) defining the molecular mechanisms of cell wall polysaccharide assembly in higher plants.

The programme of work outlined in this proposal will have considerable academic, economic and societal impact. It will not only lead to new knowledge and scientific advancement for the numerous collaborating scientists detailed in the proposal, but will benefit the wider academic community who have interests in cell-cell interactions, pathogen virulence, immune suppression, adaptive immunity, health and nutrition, the gut microbiome, human fertilization and reproduction, metabolic disorders, bioprocessing and plant sciences. Non-academic beneficiaries of our research will include the biotech and pharma industries, plus the biofuels and industrial biotechnology sectors. Other beneficiaries are the Animal Health and Veterinary Laboratories Agency, who will exploit our research outputs to improve their assays for diagnosis of porcine and bovine brucellosis, and the Defence Science and Technology Laboratory, whose vaccine programmes will benefit from our enhanced methodologies for bacterial LPS analysis. Many of the BBSRC's priority areas have an urgent need for glycobiological skills. We will address this issue by training and delivering highly skilled glycobiology researchers in our GlycoTRIC Centre. We will continue to develop and improve glycomic and glycoproteomic technologies, especially our glycoinformatic tools and data depositories, which will lead to worldwide academic advancements. Our research will give outputs that will benefit and impact on society in general. For example our studies of the glycobiology of host/pathogen interactions will lead to the development of new treatments and vaccines against a wide range of human and animal diseases which will lead to better human and animal health and an improved quality of life and food security. Our studies of the glycobiology of mammalian fertilization and reproduction will assist women suffering from infertility and recurrent pregnancy loss and potentially also have an impact on livestock production. Our studies of plant cell wall biosynthesis will provide a better fundamental understanding of the processes involved and therefore facilitate the exploitation of biomass for the production of biofuels and high value chemicals. Outputs from our glycomic and glycoproteomic method development will have an impact on UK and international industry therefore contributing towards wealth creation and economic prosperity. Our research on defining post-translational modifications of proteins is of special interest to both the pharma and biotech industries, not least because of the importance of glycosylation in many therapeutic proteins currently under development. Our mass spectrometric method development has been pioneering, and a number of our original discoveries including the concept of MS peptide and glycopeptide mapping now feature in the International Committee on Harmonisation (ICH) Guidelines for Test Procedures and Acceptance Criteria for Biotechnology Products and in the Points to Consider advice on Characterising Monclonal Antibodies, thus accelerating and simplifying the characterisation of Biopharmaceutical Products with considerable financial implications of benefit to industry and to the end-consumer in society. Our research outputs will benefit MS instrument manufacturers in the UK and abroad. Such companies can exploit our glycoinformatics developments by incorporating the tools we design into their commercial analysis platforms, benefiting from reduced development costs and increased sales as a result of the new features gained. Bruker Daltronics recently benefited in this way by incorporating our GlycoWorkbench tool into their commercial software.