Organisms of the gut-invasive genus Helicobacter, such as H. felis shown, have long been a hallmark of BABS research and are the present focus of Professor Hazel Mitchell.
Tethered-catalysis Two-hybrid System: Identification of Modification-regulated Interactions in the Interactome
The Wilkins lab is currently focused on the construction and analysis of protein interaction networks. This is a new systems biology approach that allows the integration and interpretation of proteomic and transcriptomic experiments. Projects are available in areas that study the post-translational modification of proteins, specifically protein methylation and the way in which these modifications control protein interactions.
The tethered-catalysis two-hybrid system will be used to test if interactions of specific proteins are mediated by methylation. This approach first requires fusion proteins of a methytrannsferase and a 'bait' protein to be made. The methyltransferase will methylate the prey protein in vivo. The manner in which methylation controls interaction can then be tested by combining this bait protein with a specific 'prey' protein or library of prey proteins.
Technically, this project gives students experience in a wide range of molecular biology techniques. These include DNA manipulation, cloning and sequencing, genetic engineering and transformation of eukaryotic cells, analysis of proteins by gel electrophoresis and western blotting as well as proteomic analysis using tandem mass spectrometry. Work is done in the model organism Saccharomyces cerevisiae, which is the organism of choice for systems biology research.
A number of previous honours projects in this area have resulted in publications. These include:
Huang, K.Y., Filarsky, M., Padula, M.P., Raftery, M.J., Herbert, B.R. and Wilkins, M.R. (2009) Micropreparative fractionation of the complexome by blue native continuous elution electrophoresis. Proteomics. 9: 2494-2502. Ho E, Webber R, Wilkins MR (2008) Interactive three-dimensional visualization and contextual analysis of protein interaction networks. Journal of Proteome Research 7: 104-12.
Couttas TA, Raftery MJ, Bernardini G and Wilkins MR (2008) Immonium ion scanning for the discovery of post-translational modifications and its application to histones. Journal of Proteome Research. 7: 2632-2641.
Other relevant links are:
NSW Systems Biology Initiative, which highlights some aspects of systems biology of the group: www.systemsbiology.org.au
Interactorium demonstration on YouTube illustrates the software the group has developed to analyse protein interaction networks: http://www.youtube.com/watch?v=WTHtYZcH6fk
BABS personnel that are responsible for this project