NORSTRUCT offers an efficient pipeline from recombinant protein production, via preliminary characterization, to crystal structure determination.
Protein samples are subjected to dynamic light scattering (DLS)analysis prior to crystallisation trials in order to estimate the homogenity of the sample. Circular Dicroism and NMR facilities are available at the Department of Chemistry and MS-MALDI is available at Department of Pharmacy.
Automatic setup of crystallisation experiments is possible using an Impax crystallisation robot.
X-ray data are collected locally using an image detector system with a rotating anode generator. The lab is also frequent users of synchrotron facilities in Europe (ESRF, Bessy, SLS).
Computing facilities for structure determination and structure-function relations include a network of PCs running on linux and windows. A couple of Silicon Graphics (SGI) workstations are also available. Protein structures are routinely solved by molecular replacement, MAD and SAD methods.
Other facilities at NorStruct include microcalorimetry and biocomputing.
The lab has equipment for culturing bacterial and fungal cells, primarily E.coli and P.pastoris, in small and large scale.
Small scale fermentation and optimization is possible using an Infors HT Sixfors bioreactor (6 x 500 ml), while larger volumes (up to 10 l) are produced in the Infors Techfors S 15 L TV fermentor. Both fermentors are capable of automatically regulating temperature, pH and oxygen. The Techfors is also able to automatically control the flow of oxygen.
Proteins are routinely produced with N- or C-terminal tags, such as His, GST and MBP. Proteins where no structure exists of hologouos proteins are produced with Selenio-methionine (Se-Met) in order to overcome the crystallographic "phase problem".
| The purification lab is well equipped with chromatographic systems from GE Healthcare:|
The lab is also equipped with a range of columns for both small and great scale protein purification as well as separation material for the most commonly used separation techniques, as affinity, ion exchange, HIC and size exclusion.
The lab routinely collects X-ray data at synchrotron facilities such as the European Synchrotron Facility (ESRF) in Grenoble, Bessy in Berlin, and Swiss Light Source (SLS) in Zürich.
The laboratory is equipped with a differential scanning calorimeter (DSC) and a isothermal titration calorimeter (ITC). These are routinely used to investigated not only the thermodynamic properties of macromolecules and interactions with other molecules but also to explore the kinetics of reactions catalyzed by enzymes.
Silicon graphics and Linux workstations are used to investigate experimental and homology modeled atomic structures of macromolecules. More sophisticated computational analysis of macromolecules and their complexes are carried out using resources at the High Performance Computing facilities located at the University campus. Such investigations include molecular dynamics simulations, free energy calculations, scoring and docking problems, protein flexibility, just to mention a few examples.