Research objectives and content
Small G-proteins belonging to the ras superfamily acts as molecular switches, linking external signals to a wide range of basic biological functions. Among these, the members of the rho-family are involved in regulation of cytoskeletal formation, cell-cycle progression, cell proliferation and apoptosis. The molecular basis of the interaction between these proteins and their downstream partners does not parallel the vast amount of genetic and biochemical information available. The research objectives of this proposal are aimed to fill this gap, by determination of the structure of one of these small G-protein, rhoA, in complex with the rhoA binding domain of one its downstream targets, the ROK kinase. The work will involve the cloning of ROK constructs encompassing the rhoA binding domain, their expression and purification for biochemical and biophysical characterisation, crystallisation of the complex and the determination of the crystal structure.
Training content (objective, benefit and expected impact)
The kind of work to be done involves most of the advanced biochemical and biophysical techniques. It is expected that all these techniques will be usefully employed in Italy, in order to increase the human capital invested in structural biology, a field of life sciences largely neglected in the past, which is beginning to receive the attention it deserves.