Theory

The macromolecular crystallography beamline (formerly known as the high-throughput protein crystallography beamline) is a dedicated facility for determining the structure of protein crystals and undertaking initial assessment of more complex crystals. Single crystals can be analysed using single wavelength anomalous dispersion (SAD) or multiple wavelength anomalous dispersion (MAD).

Expert technical assistance is available to help users who are not specialist crystallographers.

Features

• high-throughput, high-resolution structure determination
• rapid, automatic data collection and interpretation to provide detailed crystal structure information in real time
• sample mounting robot (SSRL type) for robotic loading and centering of crystals
• optional manual mounting of crystals directly onto gonio
• remote robot operation
• beam monitoring
• motorised collimating slits (x2)
• fast shutter
• sample video microscope (coincident with the beam) embedded in the Blu-Ice User Interface
• COMING SOON: Phi or Kappa goniostat (interchangeable) and support

Techniques Available:

• High resolution single wavelength anomalous dispersion (SAD)
• Multiple wavelength anomalous dispersion (MAD)

Applications

The rapid determination of large numbers of protein structures is now essential in many fields of science, including molecular biology, rational drug design and proteomics.

For example, protein crystallography provides primary structural information on complex macromolecules that drive biological processes.

Proteomics involves systematic characterisation of the full set of an organism’s gene products, with a key component being structural genomics - the elucidation of three-dimensional protein structures.

X-ray diffraction is the most widely-used method for protein structure determination, providing information for a wide range of applications, including drug development, food technology, agriculture, manufacturing and chemical processing.

Research

Life sciences

• Protein structure

Physical sciences

• Chemical structure

Examples of synchrotron x-ray crystallography applications

• determination of the structures of protein complexes, providing valuable information for other synchrotron studies of mechanisms and ligand binding at atomic resolution
• structural studies of proteins critical to insect physiology to inform the development of new or improved insecticides
• structural studies of transport proteins in plant roots to improve knowledge of salt resistance mechanisms used by native plants and agricultural crops
• structural studies of malarial proteins to assist the development of drugs that block malaria parasite entry to red blood cells
• structural studies of growth factors to improve understanding of cancer growth and develop better treatments
• investigation of the global structures of large biological molecules, to complement information obtained from NMR and other studies, and investigate the regulation of key cellular processes that underpin healthy biological functioning or are implicated in disease states
• structural studies of antibodies, to assist the identification of bio-sensor platforms for cancer detection.
• determination of the structure of the anthrax lethal factor, making it possible to pursue the development of anti-toxins.

Case study 1: Rational drug design – influenza

The ability of synchrotron x-rays to reveal the detailed structures of biological proteins and their interactions has enabled researchers to develop a new approach to drug discovery. Rational drug design identifies opportunities to block or modify molecular interactions. The anti-influenza drug Relenza^TM is the world’s first structure-based anti-viral drug and an early example of rationally based drug design methodologies. Relenza^TM was developed in the mid-1990s by a CSIRO team led by Peter Coleman and Jose Varghese.

Coleman and Varghese used synchrotron protein crystallography to create a high-resolution picture of the neuraminidase protein on the virus surface.

Case study 2: Therapeutics for chronic inflammatory diseases

Chronic inflammatory diseases represent one of the greatest health problems in the developed world, and macrophages play a central role in the inflammation process. Researchers from the University of Queensland are investigating macrophage proteins from mice. They want to develop a better understanding of the inflammation process in arthritis and other chronic inflammatory diseases and to identify targets for the development of new anti-inflammatory therapeutics.

Associate Professor Jenny Martin and her colleagues have established a bacterial expression system to screen hundreds of proteins to identify those that are suitable for structural studies. Proteins that express well in the small-scale bacterial system are then produced in large scale and evaluated further by structural techniques.

Configuration Modes

In contrast to other beamlines at the Australian Synchrotron, the configuration of the Macromolecular Crystallography (High-throughput Protein Crystallography) endstation is fixed such that all user groups employ a standardised gonio head and protein crystal mounts for analysing crystals. Further detail on crystal mounts and sample environments are detailed in the Samples webpage for this beamline.

The following series of photos and illustrations outlines the optical configuration of the beamline; from the synchrotron machine wall (‘white’ beam) through to the endstation (single wavelength X-rays onto sample). Further information for beamline users can be obtained in the Technical Information webpage for this beamline.

Beamline Overview: Key Components

Download of Schematic Diagram (pdf 324 KB)

Photon Delivery and Beam Conditioning: the Machine Wall and Optics Hutch

Wavelength Selection and Beam Steering/Focusing: Inside the Optics Hutch

Beam Focusing, Sample Environment and Detectors: the Experiment Endstation

Endstation Detail: Robot and dewar (left of photo), sample environment (centre of photo), and ADSC Quantum 210r Detector (right of photo).

Beamline Controls and Sample Preparation: the User Cabin

• Blu-Ice user GUI
• Light microscopes
• Data storage array (~40TB)
• Data processing computers