UQROCX Crystallisation Facility

Remote operation crystallisation and X-ray diffraction facility

UQ ROCX provides access to world-class equipment for protein crystallisation and diffraction studies to solve crystal structures. Services include protein crystallisation condition screening and optimisation, crystal diffraction screening, data collection, data processing, and structure determination. Nano-litre liquid handlers and automated imaging means that large numbers of crystallisation conditions can be investigated with small quantities of protein.

Researchers are using the facility to draw electron density maps that resolve protein structures at atomic resolution, with applications in protein functional studies, drug design and screening fragment libraries for drug leads.

Recent highlights

  • Structures of proteins required for plant immunity against fungal pathogens
  • Studies of sorting nexins required for receptor recycling, signalling and homeostasis
  • High-resolution structures of Mycobacterium tuberculosis drug targets
  • New insights into proteins required for synaptic neurotransmission

High-resolution structural information is necessary in understanding these important processes. The research is providing crucial insights into disease-causing mutations, which is boosting the future design of drugs that affect their activity.

The diffraction facility has Queensland’s brightest research X-ray source and the state’s only robotic sample storage and retrieval system, which allows for multiple data sets to be collected without user intervention.

The facility is funded by the The University of Queensland.

Equipment available

Rigaku FR-E SuperBright X-ray generator

The Rigaku FR-E+ SuperBright is the most intense home laboratory X-ray source available today for macromolecular crystallography. Its brightness exceeds some second-generation bending magnet synchrotron beamlines.

  • Vari-Max HF optic left port
  • HiRes2 optic right port
  • CCD and image plate detectors

Rigaku Saturn 944 CCD area detector

A highly sensitive CCD detector for very fast data collection. The Rigaku Saturn 944 CCD detector can collect a protein data set in 30 minutes.  When teamed with the ACTOR crystal mounting robot, fully automated crystal screening and data collection is possible.

ACTOR™ – Automated Crystal Transfer, Orientation and Retrieval

ACTOR enables screening of up to 30 samples per hour. Adding automation to your structure solution pipeline also complements the high throughput needed for fragment-based screening projects and structural genomics programs. ACTOR enables quick screening of many crystals for optimal diffraction qualities and cryo-conditions and includes ranking software to assist sample comparison and finding the best crystals for data collection.

Rock Imager

Rock Imager is an automated imaging system for protein crystallisation. This robust, easy-to-use solution incubates and captures superior-quality images of up to 1000 microplates on a user-defined schedule. Load your plates and go.

  • Superior optics and intelligent imaging provide industry-leading image quality
  • Automatically images plates on user-defined schedules
  • Peltier heat exchanger regulates temperature to +/- 0.5 °C with a range of 5 °C below or 7 °C above room temperature
  • Compressor cooling option (RI1000 only) for precise temperature control from 30 °C down to 4 °C
  • SBS and Microbatch plate compatible (Linbro and Qiagen EasyXtal plates optional)
  • Seamless integration with Rock Maker or your current crystallization software via Rock Imager API

Tecan liquid handling robot

Used to custom formulate crystallisation condition solutions. Compact liquid handling power for small life science laboratories in academia, biotechnology research and analytics or as a personal system for optimising processes independently in larger pharmaceutical or agrochemical companies. The freely configurable Freedom EVO 75 supports applications such as DNA extraction, amplification set up, sample dilution, normalisation and assay development. An optional robotic manipulator arm, also able to reach positions outside the worktable, enhances the system to provide a complete process automation platform in a small footprint.

Mosquito Crystal & Mosquito LCP

Mosquito is the ideal liquid handling tool for setting up protein crystallisation drops in high-throughput screening plates. Vapour diffusion and LCP methods are supported. Mosquito offers solutions and protocols for sitting drop or microbatch in all commercially available multi-well plates and provides a solution for hanging drop on a 96 well format.

Rigaku R-Axis IV++ area detector

R-Axis IV++ area detector has very high dynamic range and can collect very high resolution data, although it is not as fast as the Saturn 944.

Viscotek TDA 305

Triple detectors are capable of measuring concentration, molecular weight, intrinsic viscosity and molecular size Rh and Rg of natural and synthetic polymers and proteins. Applications include studying molecular weight, aggregation, conjugates and copolymers, and molecular structure.

Crystallisation Screen Conditions

Available commercial screening solutions

  1. Index: This plate contains Hampton Research Index Screen (HR2-144)
  2. JCSG: This plate contains Molecular Dimensions JCSG-plus Screen (MD1-37)
  3. ShotGun: This plate contains Molecular Dimensions SG1 Screen (MD 1-88)
  4. PACT: This plate contains Molecular Dimensions PACT premier Screen (MD1-29)
  5. Proplex: This plate contains Molecular Dimensions ProPlex Screen (MD1-38)
  6. PEG Ion: This plate contains Hampton Research
    PEG/Ion (HR2-126) Rows A-D
    PEG/Ion 2 (HR2-098). Rows E-H
  7. PEGRX: This plate contains Hampton Research
    PEGRx 1 (HR2-082) Rows A-D
    PEGRx2 (HR2-084). Rows E-H
  8. Combined (Combo): This plate contains Screen Hampton Research Grid Screens
    Sodium Malonate (HR2-247) Rows A-B
    Ammonium Sulfate (HR2-211) Rows C-D
    Sodium Chloride (HR2-219) Rows E-F 
    PEG/LiCl (HR2-217) Rows G-H

Services

Protein crystallisation

Screening for conditions suitable for crystal growth and the optimisation of these conditions to produce diffraction quality crystals is performed with the aid of commercial conditions and low-volume liquid handlers.

Commercial screening conditions and/or custom screening is performed using the Tecan and Mosquito liquid handler. Screening plates are monitored in the RockImagers. Optimised conditions are prepared using the RockMaker software and the Tecan liquid handler.

X-ray safety training

Government regulations require all users to have a licence under the Radiation Safety Act 1999.  This training combined with the Analytical X-ray safety course run by UQ OH&S is the starting point to becoming licensed.

Training to become a licensed user

Full training requirements to becoming a licensed X-ray user can be provided at an hourly rate; the time taken varies from potential licensee to licensee. For someone with no previous X-ray experience a minimum of 15 hours would be needed.

Full data collection

We can offer a full service, from crystal growth to data collection and solution and refinement. Experienced crystallographers are available to crystallise, mount and collect your data. Data can also be processed in house, or raw data images supplied. Charges are available upon request.

Further details of this facility and its available services can be found at http://uqrocx.imb.uq.edu.au.