Novel tools for crystallisation of macromolecules
FP7-PEOPLE-IAPP -Industry-Academia Partnerships and Pathways. EU funding: 514 000 €. in collaboration with Imperial College, UK and FARFIELD, Inc., UK. 1/3/2008 to 29/2/2012. E. Saridakis, PI and Coordinator.
Knowledge of the structure of biological macromolecules, especially proteins, at the molecular level, is necessary to the understanding of the biochemical mechanisms on which life depends, but also to key biomedical applications, such as rational drug design. It is also crucial to the chemical industry, agricultural research and bionanotechnology. X-ray crystallography is the most widely used method that allows the elucidation of the structure of large macromolecules at high, even atomic (1 Å and sometimes better) resolution, i.e. the atom-by-atom determination of their three-dimensional structures.
In the long process of the structure determination of a biological macromolecule, crystallisation is considered to be the most difficult and intractable step. Programme TOPCRYST (Tools for Protein Crystallisation) developed new methodology which allows the fast and effective determination of conditions for the crystallisation of any given protein. We have found that a technique known as Dual Polarisation Interferometry (DPI) allows to observe early-stage crystal nucleation in a protein solution during a dialysis experiment and to distinguish this from the alternatives states of either fully soluble protein or amorphous aggregation. This feature will allow the determination of crystallisation conditions, complementing the standard time-consuming and unreliable method of trying hundreds of potential conditions until hitting a correct one. TOPCRYST was a four-year Programme under FP7 category PEOPLE, sub-division “Industry-Academia Partnerships and Pathways”. It therefore centrally involved the exchange of technical expertise between an SME based in the U.K., Farfield Group Ltd. and the N.C.S.R. “DEMOKRITOS”, through the mutual temporary secondment of staff. A third partner was the laboratory of biological crystallisation (Prof. N. Chayen) of the Faculty of Medicine, Imperial College London. Farfield has developed and is producing the DPI instrumentation whereas the Laboratory of Structural and Macromolecular Chemistry of the Institute of Physical Chemistry of DEMOKRITOS (Drs. E. Saridakis, I. Mavridis, Κ. Yannakopoulou) and Imperial College provided expertise in biological crystallisation as well as the proteins that are used in the project.