The Netherlands Cancer Institute (NKI) is an integrated hospital and research centre. The NKI research division houses about thirty basic research groups, in eight departments. The Department of Biochemistry houses three research groups, including that of Anastassis Perrakis and the Protein Facility: both are key contributors to this project. The Protein Facility has extensive expertise and equipment for expression of proteins in E. coli, insect cells and mammalian cells and uses several protein purification stations. It also operates a nano-crystallization robotics facility, including robotics for screening (vapour diffusion and microfluidics) and for optimization, and two fully automated imaging stations (at room temperature and at 4 oC). These, together with equipment for protein quality control (a multi-angle laser light scattering device (MALLS) from Wyatt with refractive index and UV measurement coupled to an FPLC from GE Healthcare, OPTIM 1000 for denaturation and aggregation characterization; Lab901 for purity control and an Orbitrap XLQ MS/MS for protein and impurities identification) will be key to provide the “Structural Audit” Access mode to new users. At the same time, the Protein Facility makes available a large variety of equipment for biophysical analysis of macromolecular interactions, including surface plasmon resonance (BIACORE T200), isothermal titration calorimetry (VP-ITC, MICROCAL), a multifunctional plate reader (Pherastar BMG) and stopped flow (TgK Scientific) both with absorption and fluorescence (FI, FP, FRET) readout, and thermal mobility shift (Nanotemper). These will be key in providing access to biophysics for validating structural hypotheses, and integrating with the imaging methods in cells (EMBL-HD), to provide joint access in the final year of the project.
Click here for the research group of Anastassis Perrakis at the Netherlands Cancer Institute and see below the people involved in iNEXT: