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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
Phloem unloading of sucrose: cloning, functional characterisation and regulation of novel membrane transporters. Sucrose is the principal form in which plant biomass, produced in photosynthetic leaves, is transported to non-photosynthetic organs for growth and storage. Sucrose transport proteins play pivotal roles in facilitating sucrose transport around plants. Hence activities of sucrose transporters directly impact on plant growth rates and crop yields. Our aim is to isolate hitherto unkno ....Phloem unloading of sucrose: cloning, functional characterisation and regulation of novel membrane transporters. Sucrose is the principal form in which plant biomass, produced in photosynthetic leaves, is transported to non-photosynthetic organs for growth and storage. Sucrose transport proteins play pivotal roles in facilitating sucrose transport around plants. Hence activities of sucrose transporters directly impact on plant growth rates and crop yields. Our aim is to isolate hitherto unknown membrane proteins that move sucrose at high rates between cells and discover their transport properties. Expected outcomes are to better understand mechanisms and regulation of sucrose transport and hence provide novel opportunities to enhance crop yield. The project will foster a productive international collaboration.Read moreRead less
In situ measurements of the electrostatic properties inside photosynthetic reaction centres: correlation with the energy conversion function of the protein. The photochemical reaction centre is a key protein complex involved in energy conversion. It converts solar energy into chemical energy as a transmembrane charge separation. Coupling of electron and proton transfer is catalysed at the level of a ubiquinone cofactor. In order to understand how the redox properties of this cofactor are fine tu ....In situ measurements of the electrostatic properties inside photosynthetic reaction centres: correlation with the energy conversion function of the protein. The photochemical reaction centre is a key protein complex involved in energy conversion. It converts solar energy into chemical energy as a transmembrane charge separation. Coupling of electron and proton transfer is catalysed at the level of a ubiquinone cofactor. In order to understand how the redox properties of this cofactor are fine tuned by the protein environment, we plan to probe the ubiquinone site using a voltage-sensitive fluorescent dye. This exciting multidisciplinary project will contribute to the understanding of how protein matrices influence and govern the midpoint redox potential of their cofactors and the environments of theirRead moreRead less
Molecular microscopy: protein and membrane dynamics in resting and activated T cells. The aim of this research, to understand the molecular organization and dynamics of the plasma membrane that underlie the signal transduction events, is at the very heart of understanding cell communication. T cell recognition and activation initiates an adaptive immune response to invading pathogens and structurally altered proteins that can be found in cancers. By providing functional insights into the molecul ....Molecular microscopy: protein and membrane dynamics in resting and activated T cells. The aim of this research, to understand the molecular organization and dynamics of the plasma membrane that underlie the signal transduction events, is at the very heart of understanding cell communication. T cell recognition and activation initiates an adaptive immune response to invading pathogens and structurally altered proteins that can be found in cancers. By providing functional insights into the molecular mechanism of T cell activation, we will not only provide fundamental knowledge of receptor signalling but also specific details of T cell receptort triggering that may lead to the development of new therapeutic strategies to control T cell activation.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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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
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
In vivo studies on the regulation of transcription and translation in Bacillus subtilis. Genetic information from a chromosome made of DNA is converted (transcription) into RNA that is then converted (translation) into protein. Transcription and translation are both highly regulated processes, and in bacteria are thought to occur very close together. Recent evidence suggests this close coupling is not as extensive as thought. This project will continue work to investigate the composition of tran ....In vivo studies on the regulation of transcription and translation in Bacillus subtilis. Genetic information from a chromosome made of DNA is converted (transcription) into RNA that is then converted (translation) into protein. Transcription and translation are both highly regulated processes, and in bacteria are thought to occur very close together. Recent evidence suggests this close coupling is not as extensive as thought. This project will continue work to investigate the composition of transcription complexes involved in production of different types of RNA, and also to investigate the level of translational coupling in live cells. This will provide useful information on these fundamental processes.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