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Lessons from the genome of Escherichia coli: understanding heterologous expression of eukaryotic internal membrane proteins in bacterial cells (BioIMPrOve)
Start date: Dec 1, 2012, End date: Nov 30, 2014 PROJECT  FINISHED 

Eukaryotic cells express integral membrane proteins (IMP) that exchange substances with their environment or receive signals in the form of small molecules, peptides or proteins. Most human IMPs are inherently difficult to study, because of complications in obtaining adequate amounts of stable and functional proteins. G protein-coupled receptors (GPCRs) are the largest IMP family in the human genome and constitute the most important class of drug targets in pharmaceutical discovery.Escherichia coli is the most used host for heterologous protein expressions, although yields and quality for GPCRs are usually not sufficient for protein characterizations. Currently, our comprehension of processes that influence the biogenesis of IMPs in bacteria is limited.Therefore, the goal of this proposal is to study how E. coli genes could regulate the heterologous production of eukaryotic IMPs, using GPCRs as a model system.In order to achieve this, we will make use of a selection strategy developed in the host laboratory that allows detecting functional GPCR expressed on E. coli and enriching cells for their functional expression level. We will apply this together with systematic libraries of recombined single-gene deletions or of overexpressed genes of E. coli.The information that will potentially emerge will allow a deeper understanding of the processes of heterologous expression of IMPs in bacteria and will also generate bacterial hosts optimized for enhanced GPCR production. The general results arising from this project could ultimately provide valuable information for the discovery of new drugs for therapy.With this proposal, an experienced researcher from Argentina will undertake mobility and carry out a period of: transfer of knowledge, skills diversification and career development in Europe. This will allow create long-term collaborations and the enhancement of mutually-beneficial cooperative networks relationships between the EU and his home country.

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