Rossjohn Laboratory

Understanding immune function and dysfunction.

The laboratory is currently investigating two broad, yet interrelated areas addressing pivotal molecular interactions in immunity: Our program is inter-linked to create a complete systematic study, namely host recognition, responses developed by the pathogen, and drug design to modulate and/or counteract these events.

Immunity

Here we aim to provide a fundamental advancement of knowledge of events that are central to innate and adaptive immunity. Understanding the structural and biophysical basis of MHC-restriction, TCR engagement, the structural correlates of T-cell signalling is significant; they represent central questions in the field of adaptive immunity. Moreover, investigating the structural basis of T-cell allorecognition, and T-cell mediated autoimmunity, will collectively provide clear insights into immune dysfunction. In addition, focussing on generic components of innate immunity is important, as the mechanisms underlying innate recognition, is simply unknown.

Infrastructure

The Rossjohn lab currently has the following infrastructure.
The Protein Crystallography Unit was established at Monash University in 2002.
The Unit comprises a Crystallization and X-ray diffraction laboratory, and each research group that uses the core infrastructure has their own laboratory for the expression and purification of proteins. All crystallographic-related computing is processed via a Mac-cluster
The Unit was generously funded by the following organisations, and their support was very much appreciated:

The Ian Potter Foundation
The National Health & Medical Research Council
The Victorian State Government
Monash University

The Crystallization Laboratory

The crystallization room is temperature-controlled, with the crystallization trays being stored on anti-vibration shelving. The laboratory houses three Leica stereomicroscopes and a Crystallization robot.

The X-ray diffraction laboratory

The X-ray diffraction laboratory abuts the crystallization laboratory. All in-house X-ray measurements are made using on of two Rikagu RU-3HBR rotating anode generator with helium purged OSMIC focussing mirrors as an X-ray source. One of the generators is also equipped for the use of a chromium anode. Data are collected using an R-AXIS IV++ detector. Crystals are routinely flash frozen (to a temperature of 100K) prior to data collection using the inverse phi geometry with an Oxford cryosystem

In addition, the laboratory frequently makes visits to synchrotron facilities in the United States and Australia, where more intense X-rays are available.
Advanced Photon Source { http://www.aps.anl.gov}

Monash Molecular Crystallisation Facility (MMCF)

For biophysical measurements of protein-protein interactions, the laboratory also possesses a Biacore 3000, and also has routine access to an isothermal titration calorimeter.

Within the Rossjohn lab., proteins are regularly expressed in prokaryotic and eukaryotic expression systems, and accordingly possess the infrastructure to achieve this. We also possess a wide range of FPLCs to purify the proteins. The Protein Crystallography Unit also has access to the Protein Production Unit.

In 2009 there was a significant upgrade to the X-ray and Crystallization infrastructure. These upgrades are funded by Monash University, State and Federal Governments, Australian Regenerative Medicine Institute (ARMI) and the Australian Research Council.

Australian synchrotron