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Monolayer crystallization of membrane proteins. Membrane proteins comprise 25-40% of all proteins and conduct a myriad of finely tuned reactions in every cell. Despite their importance and diversity only ~40 membrane protein structures have been solved, due to the difficulty of producing high quality 2D and 3D crystals. We propose to develop and use the new monolayer crystallization technique, which employs a lipid monolayer as a crystallization template for 2D crystal production. A number of ....Monolayer crystallization of membrane proteins. Membrane proteins comprise 25-40% of all proteins and conduct a myriad of finely tuned reactions in every cell. Despite their importance and diversity only ~40 membrane protein structures have been solved, due to the difficulty of producing high quality 2D and 3D crystals. We propose to develop and use the new monolayer crystallization technique, which employs a lipid monolayer as a crystallization template for 2D crystal production. A number of important membrane proteins are available for these structural studies including ABC transporters, Caveolin-3 and the NS1 protein of Dengue virus, all of which are difficult to crystallize using conventional techniques.Read moreRead less
Novel approaches for structural and functional analysis of the protein complex COG, a tether that controls intra-Golgi trafficking. Production and engineering of proteins are key methodologies of the life sciences in general and biotechnology in particular. Our ability to produce and analyse protein-based components of the cell determines the expense and speed of discovery, as well as the creation of new vaccines, drugs, and diagnostic methods. The current project aims to develop new approaches ....Novel approaches for structural and functional analysis of the protein complex COG, a tether that controls intra-Golgi trafficking. Production and engineering of proteins are key methodologies of the life sciences in general and biotechnology in particular. Our ability to produce and analyse protein-based components of the cell determines the expense and speed of discovery, as well as the creation of new vaccines, drugs, and diagnostic methods. The current project aims to develop new approaches for protein production and to apply them to the analysis of the basic mechanisms of cell self-maintenance.Read moreRead less
NmlR-dependent thiol-based redox systems and their role in global stress responses in bacteria. All cells sense changes to their environment and respond by altering their metabolism. A major environmental change is oxidative stress which damages cells. Cells have the ability to sense oxidative stress and alter metabolic processes to defend against the damage that it elicits. This proposal will characterize a novel oxidative stress defense system that is found in a number of bacterial pathogens t ....NmlR-dependent thiol-based redox systems and their role in global stress responses in bacteria. All cells sense changes to their environment and respond by altering their metabolism. A major environmental change is oxidative stress which damages cells. Cells have the ability to sense oxidative stress and alter metabolic processes to defend against the damage that it elicits. This proposal will characterize a novel oxidative stress defense system that is found in a number of bacterial pathogens that need to defend themselves against attack by the host. The project may identify new ways to manage these bacterial pathogens. It may also provide an insight into oxidative stress defense processes that are linked to pathologies in humans.Read moreRead less
Innovative Approaches to Membrane Protein Crystallography & Drug Discovery. Membrane proteins make up around 30% of the predicted products from our human genome, are critical for life, and represent the targets of biological agents like hormones and toxins as well as most drugs. Yet these proteins have persistently defied our best efforts to study them: we know very little about what they do or what they look like. This project is aimed at cracking the problem of membrane proteins, while at the ....Innovative Approaches to Membrane Protein Crystallography & Drug Discovery. Membrane proteins make up around 30% of the predicted products from our human genome, are critical for life, and represent the targets of biological agents like hormones and toxins as well as most drugs. Yet these proteins have persistently defied our best efforts to study them: we know very little about what they do or what they look like. This project is aimed at cracking the problem of membrane proteins, while at the same time developing screening methods that can be used to design drugs against them. The long-term benefits to the community will include fundamental new knowledge and the development of new technologies and pharmaceuticals.Read moreRead less
Subunit stoichiometry and arrangement in the glycine receptor. Glycine receptors are important for nervous system function. These receptors comprise a mixture of 5 alpha and beta subunits arranged around a central ion-conducting pore. The subunit stoichiometry (i.e., numbers of alpha and beta subunits) and arrangement (i.e., subunit order) are unknown. The first aim of this project is to define these parameters using tethered subunits. The second aim is to use the tethered subunits to probe th ....Subunit stoichiometry and arrangement in the glycine receptor. Glycine receptors are important for nervous system function. These receptors comprise a mixture of 5 alpha and beta subunits arranged around a central ion-conducting pore. The subunit stoichiometry (i.e., numbers of alpha and beta subunits) and arrangement (i.e., subunit order) are unknown. The first aim of this project is to define these parameters using tethered subunits. The second aim is to use the tethered subunits to probe the structure and function of glycine and zinc binding sites at an unprecedented level of resolution. The results will provide crucial new information concerning glycine receptor structure and function.Read moreRead less
Development of a novel high yield cell-free protein expression system. Recombinant proteins are used as vaccines, drugs, and research tools, as well as food and detergent additives, comprising a A$100 billion international market. Their production requires laborious, expensive, and time-consuming construction of transgenic organisms or cells. Alternatively, recombinant proteins can be produced in extracts prepared from cells or organisms. The aim of this proposal is to develop a new technology t ....Development of a novel high yield cell-free protein expression system. Recombinant proteins are used as vaccines, drugs, and research tools, as well as food and detergent additives, comprising a A$100 billion international market. Their production requires laborious, expensive, and time-consuming construction of transgenic organisms or cells. Alternatively, recombinant proteins can be produced in extracts prepared from cells or organisms. The aim of this proposal is to develop a new technology that will make cell-free production of recombinant proteins rapid, cheap, and scalable. This will advance Australia’s intellectual leadership in the area of biotechnology and will bring numerous economic benefits by accelerating pharmaceutical development. Read moreRead less
Evolution of halophytes: a phyloinformatic approach to understanding and exploiting the traits underlying salt-tolerance in plants. Salinity is an increasing burden on the Australian economy & environment, with >2 million ha of salt-affected land, at an annual cost to agriculture over $187 million. One solution is to exploit naturally salt-tolerant plants to increase productive agricultural land and restore salt-affected environments. To do this, we must increase basic knowledge of the diversity ....Evolution of halophytes: a phyloinformatic approach to understanding and exploiting the traits underlying salt-tolerance in plants. Salinity is an increasing burden on the Australian economy & environment, with >2 million ha of salt-affected land, at an annual cost to agriculture over $187 million. One solution is to exploit naturally salt-tolerant plants to increase productive agricultural land and restore salt-affected environments. To do this, we must increase basic knowledge of the diversity & distribution of salt-tolerance. This project is the first to use DNA sequences from thousands of species to understand the evolution of salt-tolerance in order to provide the foundation for the development of new crop varieties, selection of species that can be developed for bioremediation, and identification of traits that will be profitable targets for breeding programs. Read moreRead less
Assessing the physiological roles of ubiquitination in regulating neuronal ion channels, receptors and transporters. Significant alterations in the activity neuronal transporters and receptors occur during tissue injury and regeneration as well as in many neurodegenerative disease states. Modulation of the pathways that control these transporters is an emerging therapeutic target, however, the molecular basis of these control mechanisms remain poorly understood. The outcome of this project will ....Assessing the physiological roles of ubiquitination in regulating neuronal ion channels, receptors and transporters. Significant alterations in the activity neuronal transporters and receptors occur during tissue injury and regeneration as well as in many neurodegenerative disease states. Modulation of the pathways that control these transporters is an emerging therapeutic target, however, the molecular basis of these control mechanisms remain poorly understood. The outcome of this project will be a thorough characterisation of a novel regulatory paradigm in neurons that is likely to be crucial for neuronal development and regeneration, and will potentially provide novel therapeutic targets for various neuronal diseases.Read moreRead less
G-protein coupled receptor-mediated calcium signalling in parasympathetic neurons. External chemical stimuli act on specific cell-surface receptors of neurons resulting in an increase in the intracellular calcium ion concentration which acts as a second messenger to alter neuronal excitability. There are, however, many receptors acting through a number of closely related proteins involving complex intracellular signalling pathways which remain poorly understood. This project uses molecular, elec ....G-protein coupled receptor-mediated calcium signalling in parasympathetic neurons. External chemical stimuli act on specific cell-surface receptors of neurons resulting in an increase in the intracellular calcium ion concentration which acts as a second messenger to alter neuronal excitability. There are, however, many receptors acting through a number of closely related proteins involving complex intracellular signalling pathways which remain poorly understood. This project uses molecular, electrical and fluorescence techniques to elucidate the molecular basis for these interactions by identifying the roles individual proteins play in integrating diverse extracellular stimuli and neuronal excitablility in the peripheral nervous system.Read moreRead less
Co-divergence or opportunism: the evolution of trematode parasitism in the sea. This proposal is for fundamental research into how a major group of parasites (trematodes) has evolved in interaction with its hosts. The work is mainly the kind of 'basic science' that underpins other science without having intended immediate community benefit. However, the work depends strongly on the important task of developing better knowledge of trematodes in Australian native animals. Some of these parasite ....Co-divergence or opportunism: the evolution of trematode parasitism in the sea. This proposal is for fundamental research into how a major group of parasites (trematodes) has evolved in interaction with its hosts. The work is mainly the kind of 'basic science' that underpins other science without having intended immediate community benefit. However, the work depends strongly on the important task of developing better knowledge of trematodes in Australian native animals. Some of these parasites are pathogens of bivalves (scallops, giant clams and oysters) but almost nothing is known about them here. Another benefit of the study is in the training of several postgraduate students who will be able to contribute to the further study and management of parasites in Australia.Read moreRead less