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Structural Chemistry at the Eskitis Institute
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Structural Chemistry complements the research portfolio of Eskitis by providing structural information
in order to understand protein mechanisms at the molecular level. To improve the knowledge of the molecular
mechanisms of diseases as well as to identify possible sites of interference, one needs to have a solid
understanding of the structure and functions of proteins, their interactions with ligands, target proteins
and membranes.
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Eskitis Structural Chemistry, with protein crystallography on the one hand, and a variety of biophysical
methods on the other, provides data about structure, shape and properties of proteins and establishes
structure-function relationships to obtain a rational for future therapeutic applications.
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Through collaborative links with other groups and programs within the Institute and other Research Centres,
we also provide expertise and support to other projects under the principal research themes of Structural
Biology and Biophysics.
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Expertise
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The group has expertise in and access to a broad range of methods from molecular cloning, protein expression and purification, protein crystallography, as well as biophysical methods such as UV/Vis, circular dichroism and fluorescence spectroscopy, multi-angle light scattering, X-ray and neutron scattering, isothermal titration calorimetry, membrane and monolayer adsorption. We also maintain a well-established network of collaborations with specialist experts and project collaborators.
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Instrumentation
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Circular Dichroism Spectrometer
JASCO J-715 ORD/CD spectropolarimeter with Peltier temperature control accessory.
CD spectroscopy is used to elucidate the conformation and secondary structure, as well as the stability of proteins.
The instrument uses the JASCO software SpectraManager for data acquisition. We have developed software for further
processing of spectra and secondary structure prediction (ACDP).
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Computing
At the centre of the Structural Computation Platform (SCP) are three independent custom-built netservers with Intel and AMD QuadCore CPUs, 1 TB RAID storage, and central authentication server. All data drives on the SCP as well as the instrumentation computers are on automated tape backup. The main operating system is Linux. We maintain a suite of up-to-date crystallographic, structural and bioinformatic software. A custom-built Laboratory Information Management System (LIMS) is used for data archiving.
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Crystallisation Room
For crystallisation, we have a dedicated crystallisation room which is temperature-controlled at 16°C.
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Differential Scanning Fluorimetry (DSF)
Thermal protein unfolding can be monitored by DSF in a multi-well format. These assays are used for compounds and fragment screening. For DSF, we are using a Roche LightCycler 480 II real-time PCR system.
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Incubator shaker
Two refrigerated Innova 44R shaker incubators from New Brunswick Scientific are used for large scale expression cell cultures.
One of the incubators was donated by the Rebecca L Cooper Foundation.
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Langmuir surface film balance
Lipid monolayers are established models for lipid membranes and a useful tool in assessing protein-membrane interactions.
Monolayer measurements are done on a Langmuir surface film balance that monitors the pressure on a lipid layer at the
air-water interface. Our lab has a NIMA Model 301A for monolayer experiments.
The instrument was donated by the Rebecca L Cooper Foundation.
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Molecular Graphics
Within the X-ray suite, we have a molecular graphics room with dedicated graphics workstations for protein structure modelling and refinement.
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Multi-angle light scattering (SEC-MALS)
Multi-angle light scattering enables determination of the molecular mass of molecules from first principles. For SEC-MALS, we
have a Wyatt miniDawn Treos multi-angle laser light scattering unit and a Shimadzu RID-10 refractive index detector coupled
to size exclusion chromatography that is performed using a BioRad DuoFlow HPLC system.
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Stereomicroscope
The Leica MZ16A stereomicroscope with motor focus is used for monitoring crystallisation as well as crystal handling
and mounting.
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Stereomicroscope
A Leica MZ16 stereomicroscope with digital camera is used for monitoring and documentation of crystallisation.
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X-ray diffractometer
Within the Structural Chemistry X-ray Facility we have a Rigaku MicroMax-007HF generator with VariMax optics delivering
a beam brightness of 3.8 * 109 X-ray/sec/mm2/mR2 and a beam size of about 0.2 mm at
the sample. The instrument is equipped with an R-Axis IV++ detector and the X-stream cryo system.
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