Whisstock Lab research
About Professor James Whisstock
Professor James Whisstock is based at the Monash Biomedicine Discovery Institute Monash University where he is currently an Australian Research Council Laureate Fellow, an Honorary National Health and Medical Research Council Senior Principal Research Fellow and Scientific Director of the Australian Research Council Centre of Excellence in Advanced Molecular Imaging.
Previously, James held an ARC Federation Fellowship, he led the NHMRC Program Grant in Protease Systems Biology (2008-2012) and he was a chief investigator on the ARC Centre of Excellence in Structural and Functional Microbial Genomics (2005-2013). In 2006 he was awarded the Science Ministers Prize for life science, in 2008 the Health Minister's Award for Excellence in Health and Medical Research and in 2010 the Australian Academy of Science Gottschalk medal.
James completed his degree at Cambridge University. During his PhD (also at Cambridge) he trained in bioinformatics and in structural biology. He came to Australia as a research fellow at Monash University in January 1997 where he established his laboratory.
- Understanding the structure, function and biology of proteases and protease inhibitors
- Understanding pore forming immune effectors in immunity and developmental biology
Visit Professor Whisstock’s Monash research profile to see a full listing of current projects.
The Whisstock laboratory is interested in three mammalian proteins – Complement component-9 (C9), Macrophage Expressed Gene-1 (MPEG-1) and Perforin. These three molecules play key roles in the destruction of pathogenic microbes, virally infected cells and malignant cells. Concomitant with working on these protein complexes, we are also developing new and better approaches for sample preparation for cryogenic Electron Microscopy experiments and for determining protein structures in the context of intact, cryogenically preserved cells.
The laboratory recently reported the cryo-EM structure of the novel perforin like protein MPEG-1. This molecule is deployed by macrophages within phagolysosomes in order to destroy engulfed microbial targets. (Pang et al., Nature Communications 2019). The structure represents the first atomic resolution structure of any perforin-like protein in a bona fide pre-pore intermediate assembly. They also showed that MPEG-1 pre-pores are activated to form pores only at acidic pH, suggesting that acidification of the phagolysosome represents a crucial trigger for MPEG-1 function.
The cryo-EM structure of the acid activatable pore-forming immune effector Macrophage-expressed gene 1. Nature Communications, Vol 10, 4288 (2019)
Pang SS, Bayly-Jones C, Radjainia M, Spicer BA, Law RHP, Hodel AW, Parsons ES, Ekkel SM, Conroy PJ, Ramm G, Venugopal H, Bird PI, Hoogenboom BW, Voskoboinik I, Gambin Y, Sierecki E, Dunstone MA, Whisstock JC.
The structure further revealed that MPEG-1 uses an N-terminal multi-vesicular body of 12 kDa (MVB12)-associated β-prism (MABP) domain in order to bind membranes. However, unexpectedly, the MABP domain was positioned such that the membrane binding face pointed in the opposite orientation to other perforin-like proteins. Collectively, together with liposome binding data the data suggest that this unusual arrangement of domains permits MPEG-1 to bind to the inner leaflet of the host membrane, with pore formation into pathogenic bacteria taking place in trans. This mechanism would prevent auto-lysis of the macrophage vesicle, an event that would likely lead to macrophage destruction.
Depicting the proposed mechanism by which MPEG1 acts to kill bacteria within the macrophage phagolysosome. MPEG1 orientation prevents damage to the macrophage membrane. Pang et al., Nature Communications, Vol 10, 4288 (2019).
With respect to imaging technologies the team has been working to develop new approaches to streamline the in situ structural biology workflow. This approach permits visualization of large protein complexes, such as the mega-dalton sized pores formed by perforin-like proteins, in the context of cryo-preserved cells. In 2019, it reported the development of the Photon Ion Electron microscope (PIE-scope; Gorelick et al., elife, 2019). In this instrument, we have integrated a cryo-Focused Ion Bean Scanning Electron Microscope (cryo FIBSEM) together with a light microscope. This powerful new tool permits fluorescent imaging of sample under cryogenic conditions within the cryo-chamber of a cryo-FIBSEM. This procedure avoids having to move material between several different microscopes, thus greatly reducing ice damage and further permitting rapid, fluorescently targeted FIB-driven isolation of a cryo-lamella from the desired region of a cell or organism.
A view of the FIB/SEM chamber containing showing the in-vacuum section of the PIE-scope. An objective is mounted on a high-precision motorized stage for sample focusing (LM focus drive). The light from the external arm (cyan arrow) is delivered through a glass flange and directed into the objective by a mirror
- Structural biology (including X-ray crystallography and electron microscopy)
- Molecular genetics.
It uses these techniques to address questions in the fields of infection and immunity, blood coagulation, developmental biology and cancer. Recent work includes the structure of the fibrinolytic pro-enzyme plasminogen and a series of discoveries around the mechanism of pore formation by the mammalian immune effector perforin.
- Systemic viral and bacterial infections
- Traumatic injuries
We collaborate with many scientists and research organisations around the world. Some of our more significant national and international collaborators are listed below. Click on the map to see the details for each of these collaborators (dive into specific publications and outputs by clicking on the dots).
Student research projects
The Whisstock Lab offers a variety of Honours, Masters and PhD projects for students interested in joining our group. There are also a number of short-term research opportunities available. You are encouraged to contact Professor James Whisstock regarding potential projects that align with the presented research themes.
- Structural studies of pore forming immune effectors
- In situ structural studies of the immune synapse
- New approaches for combating clotting related disorders in infection and inflammatory disease.