Selective enrichment of proteins using micro-Gradiflow technology and characterisation of proteins using LC-MS/MS. This project describes crucial areas for the technology development of protein enrichment and fractionation. While current techniques are powerful, the usefulness to identify and characterise proteins of low abundance from massively complex samples in the presence of abundant proteins is limited and requires up to mililitre volumes. We intend to develop the micro-Gradiflow technolog ....Selective enrichment of proteins using micro-Gradiflow technology and characterisation of proteins using LC-MS/MS. This project describes crucial areas for the technology development of protein enrichment and fractionation. While current techniques are powerful, the usefulness to identify and characterise proteins of low abundance from massively complex samples in the presence of abundant proteins is limited and requires up to mililitre volumes. We intend to develop the micro-Gradiflow technology in association with Gradipore as a means of protein fractionation using microlitre volumes of sample. This technology will be applicable to all areas of proteomic research and in particular to the study of cell differentiation. The outcome for Australian industry, our competitiveness both industrial and scientific, and the potential for economic advancement is tremendous.Read moreRead less
Characterising post-translational modifications in bacterial proteins. This project represents the first global attempt to characterize post-translational modifications in bacterial proteins using the tools of proteomics. Modifications to proteins are key elements in altering their function. In bacteria, modifications are important in cell-cell adhesion, signalling and triggering of the immune response. Characterisation of modified proteins and their sites of modification represents an opportuni ....Characterising post-translational modifications in bacterial proteins. This project represents the first global attempt to characterize post-translational modifications in bacterial proteins using the tools of proteomics. Modifications to proteins are key elements in altering their function. In bacteria, modifications are important in cell-cell adhesion, signalling and triggering of the immune response. Characterisation of modified proteins and their sites of modification represents an opportunity to understand how bacterial cell populations communicate in the environment, as well as aid in understanding pathogenesis in medical, veterinary and food-borne pathogens. Therefore, improved vaccine targets and therapeutics, as well as method-based products, may be generated by this project.Read moreRead less
Improving the sustainability of Australia's water resources: an effective approach for diagnosing and treating foulants on water recycling membrane filters. By determining ways to diagnose and treat fouling problems we will have effectively solved a critical problem in the water recycling industry. By creating more efficient and sustainable ways of using water, we will transform the effectiveness of a wealth of Australian industries that increasingly rely on the efficient use of water (e.g. mini ....Improving the sustainability of Australia's water resources: an effective approach for diagnosing and treating foulants on water recycling membrane filters. By determining ways to diagnose and treat fouling problems we will have effectively solved a critical problem in the water recycling industry. By creating more efficient and sustainable ways of using water, we will transform the effectiveness of a wealth of Australian industries that increasingly rely on the efficient use of water (e.g. mining, agriculture, textiles, energy). By reducing the demand for water (by effectively recycling it), and the energy requirements (by efficiently recycling it), we will increase national wealth and provide significant social and environmental benefits to all Australians.Read moreRead less
Visualising below the tip of the proteome iceberg. Proteomics attempts to understand human biology by examining the protein components of cells and tissues. Unfortunately, currently available technology only allows approximately 10% of the complexity of these cells and tissues to be concurrently investigated. This project will the physical, chemical and functional properties of protein classes for enrichment, as well as improve technologies for protein visualization, identification and character ....Visualising below the tip of the proteome iceberg. Proteomics attempts to understand human biology by examining the protein components of cells and tissues. Unfortunately, currently available technology only allows approximately 10% of the complexity of these cells and tissues to be concurrently investigated. This project will the physical, chemical and functional properties of protein classes for enrichment, as well as improve technologies for protein visualization, identification and characterization. These approaches will allow the scientific community to further mine beneath the surface of the proteomics 'iceberg'.Read moreRead less
Dissecting a hematopietic transcription factor complex. The development of mature active cells is a highly complex and coordinated process that is controlled largely by groups of interacting regulatory proteins. We are trying to understand, at a very detailed level, how a specific group of these proteins interact to regulate both normal blood cell development and the onset of childhood leukemias. Using this information we will try to develop reagents that can be used to inhibit these interaction ....Dissecting a hematopietic transcription factor complex. The development of mature active cells is a highly complex and coordinated process that is controlled largely by groups of interacting regulatory proteins. We are trying to understand, at a very detailed level, how a specific group of these proteins interact to regulate both normal blood cell development and the onset of childhood leukemias. Using this information we will try to develop reagents that can be used to inhibit these interactions and be used as lead compounds for treatments for disease.Read moreRead less
Oxidative Damage and Cell Ageing. This research will benefit Australia by providing a fundamental understanding of how cells age. This will have immediate international impact at the scientific level and will inform strategies to reduce the rate of ageing and alleviation of age-related disorders. In the longer term the research may provide commercial and social outcomes by identifying antioxidant systems that will provide a genuine benefit in reducing ageing.
Cellular Responses to Oxidative Damage: Cell Aging. The aim of this project is to identify the mechanisms by which oxidative stress and free radical damage cause cell aging. This work will make a significant contribution to our understanding of the aging process in cells by identifying the major reactive oxygen species that contribute to cell aging, which defence systems and antioxidants provide the greatest degree of protection, what damage accumulates as cells age and which genetic systems ar ....Cellular Responses to Oxidative Damage: Cell Aging. The aim of this project is to identify the mechanisms by which oxidative stress and free radical damage cause cell aging. This work will make a significant contribution to our understanding of the aging process in cells by identifying the major reactive oxygen species that contribute to cell aging, which defence systems and antioxidants provide the greatest degree of protection, what damage accumulates as cells age and which genetic systems are activated as during the process.Read moreRead less
The effect of nitrogen monoxide on intracellular iron metabolism. We discovered that the crucial signalling molecule nitrogen monoxide (NO) mediates iron (Fe) and glutathione (GSH) release by the transporter MRP1 probably as an NO-Fe-GSH complex [DR(2006) PNAS USA 103:7670-5]. During our current ARC grant we have markedly extended these findings by showing that another molecule, GST Pi and MRP1 form part of a coordinated system that stores and transports NO as complexes of Fe and GSH, markedly e ....The effect of nitrogen monoxide on intracellular iron metabolism. We discovered that the crucial signalling molecule nitrogen monoxide (NO) mediates iron (Fe) and glutathione (GSH) release by the transporter MRP1 probably as an NO-Fe-GSH complex [DR(2006) PNAS USA 103:7670-5]. During our current ARC grant we have markedly extended these findings by showing that another molecule, GST Pi and MRP1 form part of a coordinated system that stores and transports NO as complexes of Fe and GSH, markedly extending NO half-life from milliseconds to hours. This has broad implications for understanding NO activity in many processes which have major vital health implications, including tumour cell killing by macrophages and blood pressure control.Read moreRead less
The Effect of Nitrogen Monoxide on Intracellular Iron Metabolism. For the first time, we discovered that nitric oxide (NO) is actively transported from cells by a protein that is known to also transport glutathione (GSH). This is important, as NO was thought to passively diffuse from cells. Active transport overcomes the problems of diffusion which is inefficient and non-targeted. Moreover, NO is released as a complex with iron and GSH which markedly increases its half-life. These findings have ....The Effect of Nitrogen Monoxide on Intracellular Iron Metabolism. For the first time, we discovered that nitric oxide (NO) is actively transported from cells by a protein that is known to also transport glutathione (GSH). This is important, as NO was thought to passively diffuse from cells. Active transport overcomes the problems of diffusion which is inefficient and non-targeted. Moreover, NO is released as a complex with iron and GSH which markedly increases its half-life. These findings have broad implications for understanding the activity of NO in many processes which have major health implications, including tumour cell killing by macrophages, blood pressure etc.Read moreRead less
The effect of nitrogen monoxide on intracellular iron metabolism. During our current ARC grant we discovered a novel relationship between energy metabolism and NO-mediated Fe efflux and showed that glutathione (GSH) is vital for this release mechanism (DR5,6). Intriguingly, this transport process is part of the cytotoxic effector machinery of activated macrophages against tumours, and requires further elucidation. We also showed that CO affects Fe metabolism by binding to Fe, and CO may modulate ....The effect of nitrogen monoxide on intracellular iron metabolism. During our current ARC grant we discovered a novel relationship between energy metabolism and NO-mediated Fe efflux and showed that glutathione (GSH) is vital for this release mechanism (DR5,6). Intriguingly, this transport process is part of the cytotoxic effector machinery of activated macrophages against tumours, and requires further elucidation. We also showed that CO affects Fe metabolism by binding to Fe, and CO may modulate NO's function. We will:-
(1) Examine if NO-mediated Fe release results in GSH efflux
(2) Identify the mechanism of NO-mediated Fe efflux.
(3) Assess the effect of inducing haem oxygenase 1 on Fe metabolism
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