Anàlisi estructural, funcional i dinàmic de l'agregació proteica i dels cossos d'inclusió en bactèries recombinants

The main obstacle for recombinant protein production in bacteria is the aggregation of the produced polypeptides a inclusion bodies, in which the products are insoluble and inactive. This fact restricts both the eventual exploitation of industrially relevant proteins and the routine production for crystallography and proteomic studies. Inclusion bodies have been considered as dead-end products of an irreversible misfolding pathway and they have been poorly studied. The approaches to minimize their formation are based on empirical observations and in are general not very efficient. Recently, we have shown that inclusion body formation is reversible, and it is deeply integrated in the quality control system of the cells, formed by a redundant network of chaperones and proteases with a high degree of functional redundancy. The objectives of this project are the biological and structural characterization of bacterial inclusion bodies to solve the nature and mechanics of protein aggregation and disagregation in vivo, as well as the identification of the cell elements that regulate both processes. In addition, we will explore metabolic engineering of the quality control system for a maximal production of soluble protein and a lesser impact of the conformational stress on the physiology of the producing cell