Proteomics of MS
Proteomics is the study of the proteome. The proteome is all the proteins expressed in a cell. Proteomics uses techniques to simultaneously quantify and identify thousands of proteins in biological samples. These samples include cerebrospinal fluid, tissue biopsies or cell cultures. Most research to date has focussed on investigating a limited number of specific molecules and this has resulted in slow progress in our understanding of the pathogenesis of MS. This national research project aims to discover the proteins involved in the development and pathology of MS. It is the first of its kind in Australia and one of the first of its kind in the world.
The MS Research Australia Proteomics Platform
The MS Research Australia Proteomics Platform is a major national MS collaboration between three Australian universities and the Sir Charles Gairdner Hospital, with the University of Adelaide as lead institution. It has a steering committee of investigators with expertise in proteomics and associated molecular technology led by Professor Shaun McColl. The platform commenced in 2010 and was funded over four years through the Australian Research Council’s (ARC) Linkage Projects funding scheme in partnership with MS Research Australia.
This platform aimed to develop state-of-the-art proteomics technologies for the Australian MS research community and use the technology to identify specific molecular changes in the CNS proteome that correlate with different stages of disease in MS. The result of this research is likely to lead to the identification of new proteins that can be targeted for the development of new therapies and diagnostics for MS.
The MS Research Australia Proteomics Platform has established facilities that are available for use for researchers in the MS community in collaboration with proteomics investigators in Adelaide and Sydney. Please use the contacts below for further information about specific facilities.
The Adelaide Proteomics Centre
The Adelaide Proteomics Centre is located at the School of Molecular & Biomedical Science at the University of Adelaide.
The Adelaide Proteomics Centre has 7 mass spectrometry instruments including LTQ Orbitrap, MALDI-TOF/TOF and Q-TOF instrumentation. The Centre has developed highly sensitive and accurate technologies for comparative proteomics experiments using label-free or a range of labelling techniques. The Centre offers all basic proteomics services including 1D, 2D and 2D-DIGE Gel electrophoresis and western blot analysis, protein identification and identification of post translational modifications by mass spectrometry and Free-Flow Electrophoresis for preparative protein separation. The centre also offers targeted protein quantitation for proteins and peptides using selective reaction monitoring (SRM) mass spectrometry methods and has established the innovative new proteomics technology of tissue imaging mass spectrometry (IMS) that is available in only a few proteomics facilities in the world. This allows a molecular profile to be generated and matched to microscopic features within the tissue. The technique has been optimised in both fresh-frozen tissue as well as archived, formalin-fixed paraffin-embedded tissue. Alongside IMS, classical proteomics using label-free quantitation of laser micro-dissected regions of interest can be performed.
For more information about the facilities at the Adelaide Proteomics Centre please email Associate Professor Peter Hoffman.
The Sydney Proteomics Centre
The Mass Spectrometry Core Facility at the University of Sydney will be located in the Charles Perkins Centre’s research and education hub, a new 49,500 square metre state-of-the-art building designed to support collaboration and new ways of thinking. The new facility builds on the 10 years of expertise developed by the Sydney University Proteome Research Unit and as a result of a multi-million dollar investment will have excellent capabilities in: quantitative, peptide centric (or bottom up) proteomics; analysis of post-translational modifications; top-down proteomic; glycomics and metabolomics. By mid-2014 over 8 mass spectrometers will be available to the MS Research Australia Proteomics Platform together with expert advice and training. The facility has many established protocols, particularly in the area of ‘discovery proteomics’, for example the use of iTRAQ labelling in the quantitative analysis of laser guided micro-dissected formalin-fixed tissue. A relatively new area is that of ‘targeted proteomics’ which uses approaches such as selective reaction monitoring (SRM) which can be an attractive alternative to methods such as western blotting or ELISA to provide rapid quantification of a number know proteins across a large number of samples.
For more information about the facilities at the University of Sydney’s Mass Spectrometry Core Facility please email Dr Ben Crossett.
MS Research Australia Proteomic Incubator Grants
MS Research Australia offers grants for studies gathering pilot data under the Incubator Grant Scheme. Incubator grants provide seed funding for the early stages of new research efforts, with the aim of generating preliminary data needed to support future grant applications. Incubator grants are awarded throughout the year with a value up to $25,000.
The MS Proteomics Research Platform now invites interested researchers to contact them to discuss their proteomics projects and the opportunities to apply for seed or project funding in collaboration with the platform. MS Research Australia would particularly like to encourage exploratory projects through the Incubator Grant ‘Request to Submit’ mechanism. The usual criteria for Incubator grants will apply.
For more information please email Professor Shaun McColl.
Global assessment of CNS-infiltrating lymphocytes
One of the major cell types believed to control the damage to the central nervous system during MS are T lymphocytes, which accumulate in the brain and spinal cord (CNS). The team has applied the new mass spectrometry platforms to conduct the first two global assessments of the proteomes of T lymphocytes invading the CNS during MS-like disease in the mouse.
The team have identified over 300 proteins in a chronic-progressive model and almost 1500 proteins in the relapsing-remitting model of MS, many of which have not previously been implicated in MS. The team are currently conducting a deeper analysis of the data using a bioinformatics approach to narrow down the most significant molecules that warrant further investigation. Three proteins have proved to be of particular interest, which are the subject of ongoing experiments.
Excellent progress has also been made in further investigations in the individual projects of the collaborating MS researchers. Imaging mass spectrometry techniques have been optimised in samples of healthy human optic nerve and in collaboration with Professor Bill Carroll, from the Sir Charles Gairdner Hospital in Perth, human optic nerves affected by MS are now undergoing analysis.
Dr Judith Greer at the University of Queensland has been developing a new form of therapeutic agent directed at specifically suppressing the immune responses against the most abundant myelin protein in the central nervous system, myelin proteolipid protein (PLP). Dr Greer has shown that an immune response to this particular protein is associated with MS lesions in parts of the brain that control balance.
In conjunction with Associate Professor Michael Barnett and Dr Ben Crossett from the University of Sydney, the team is also analysing human MS lesions using complementary mass spectrometry techniques, including imaging mass spectrometry.
The breadth of these projects and the combined expertise of the investigators will see some very exciting progress and insights into the molecules that lie at the heart of MS pathology and will allow the development of highly targeted new treatments for MS.