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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Linkage Infrastructure, Equipment And Facilities - Grant ID: LE0882295
Funder
Australian Research Council
Funding Amount
$225,000.00
Summary
X-ray crystallography resource for membrane proteins and large macromolecular complexes. Structural biology is the underpinning of biotechnology, biopharmaceuticals and rational therapeutic design. The most successful technique for determining the structures of proteins and large macromolecular complexes is x-ray crystallography. This proposal will set up a network of state of the art resources in the Sydney region to capitalise on expertise in these areas. The facilities will foster basic re ....X-ray crystallography resource for membrane proteins and large macromolecular complexes. Structural biology is the underpinning of biotechnology, biopharmaceuticals and rational therapeutic design. The most successful technique for determining the structures of proteins and large macromolecular complexes is x-ray crystallography. This proposal will set up a network of state of the art resources in the Sydney region to capitalise on expertise in these areas. The facilities will foster basic research and collaborations with industry, which will enhance Australia's profile and commercialisation of research. The facility will enhance the usage of the Australian synchrotron, producing flagship projects on the edge of technical possibilities.Read moreRead less
How do cells regulate redox environment at the subcellular level? Most organisms live in an aerobic environment that subjects their cells to reactive oxygen species. Reactive oxygen species have been proposed to lead to ageing, and in many diseases the balance between oxidising and reducing conditions (the redox environment) is perturbed. This research will identify how different cellular structures sense and maintain this redox homeostasis, not just in the whole cell, but within the different ....How do cells regulate redox environment at the subcellular level? Most organisms live in an aerobic environment that subjects their cells to reactive oxygen species. Reactive oxygen species have been proposed to lead to ageing, and in many diseases the balance between oxidising and reducing conditions (the redox environment) is perturbed. This research will identify how different cellular structures sense and maintain this redox homeostasis, not just in the whole cell, but within the different organelles in the cell. The work will help identify which cell compartments and processes are affected in different disease states and provide a fundamental understanding of how cells coordinate their different organelles to maintain the balance between oxidising and reducing conditions.Read moreRead less
Protein methylation: a fundamental regulator of the interactome. Proteins are the functional molecules of the cell. They interact with each other to form small 'protein machines' that are part of large, complicated networks. This study will examine how the cell makes tiny changes to proteins, through the addition of one carbon and two hydrogen atoms, and how this is important in the regulation of protein interactions. The proteins of baker's yeast, a common model organism, will be studied here. ....Protein methylation: a fundamental regulator of the interactome. Proteins are the functional molecules of the cell. They interact with each other to form small 'protein machines' that are part of large, complicated networks. This study will examine how the cell makes tiny changes to proteins, through the addition of one carbon and two hydrogen atoms, and how this is important in the regulation of protein interactions. The proteins of baker's yeast, a common model organism, will be studied here. However, the findings will be directly relevant to understanding the function of many proteins in plants, animals and man.
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Investigating Post-transcriptional Gene Regulation In Cancer
Funder
National Health and Medical Research Council
Funding Amount
$645,205.00
Summary
In this program, I will enhance our understanding of cancer gene regulation and provide novel avenues for the treatment of aggressive tumours. Using own data and that from collaborators, I will determine patterns of gene regulation in blood cancers and identify markers that predict disease outcome. I aim to understand how gene regulation can transform healthy cells into tumour cells and whether personalised treatment can kill tumour cells more effectively and prevent relapse and metastasis.
Ubiquinone in Giardia: Amitochondrial component in an amitochondriate parasite. Giardia intestinalis is a fascinating organism, it is one of the most primitive nucleated organisms known and is responsible for ~280 million infections annually. Ubiquinone is usually associated with mitochondrial function, however it has been found in Giardia, which lacks this organelle. Our initial studies show that in Giardia, ubiquinone plays essential roles in electron transport pathways associated with membr ....Ubiquinone in Giardia: Amitochondrial component in an amitochondriate parasite. Giardia intestinalis is a fascinating organism, it is one of the most primitive nucleated organisms known and is responsible for ~280 million infections annually. Ubiquinone is usually associated with mitochondrial function, however it has been found in Giardia, which lacks this organelle. Our initial studies show that in Giardia, ubiquinone plays essential roles in electron transport pathways associated with membrane energisation and oxidative stress management. Elucidation of these mechanisms will have a major impact on the understanding of Giardia and other anaerobic organisms as well as being of significant evolutionary and medical importance.
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Characterisation of the CLIC1 chloride ion channel by a novel biophysical method: Site-Directed-Spin-Labeling Electron Paramagnetic Resonance Spectroscopy. Chloride ion channels are involved in diverse physiological processes and channel malfunction can lead to severe diseases. This project examines the structure and conformational changes of a member of the newly described chloride channel family (CLIC1) using an emerging biophysical technique. CLIC1 is unique due to its ability to transit be ....Characterisation of the CLIC1 chloride ion channel by a novel biophysical method: Site-Directed-Spin-Labeling Electron Paramagnetic Resonance Spectroscopy. Chloride ion channels are involved in diverse physiological processes and channel malfunction can lead to severe diseases. This project examines the structure and conformational changes of a member of the newly described chloride channel family (CLIC1) using an emerging biophysical technique. CLIC1 is unique due to its ability to transit between soluble and active membrane channel forms. Our novel approach to determine the channel structure represents a major advance in overcoming numerous difficulties associated with traditional atomic resolution structural-biology techniques. This proposal also opens up new experimental avenues to understand biological important events associated with ion channels, including channel gating.Read moreRead less
A unified model of amino acid homeostasis. This project aims to develop a unified model of amino acid homeostasis in mammalian cells and apply it to brain cells. The model will be underpinned by a mathematical algorithm that allows predicting amino acid levels in the cytosol based on fundamental parameters such as transport and metabolism. This project should provide the significant benefit of enabling the prediction of essential functions such as cell growth and survival.