GPCR EVOLUTION

Directed evolution of class B G-protein coupled receptors

Coordinatore	UNIVERSITAET ZUERICH    more  Organization address address: Raemistrasse 71 city: ZURICH postcode: 8006 contact info Titolo: Prof. Nome: Andreas Cognome: Plückthun Email: send email Telefono: 41446355571 Fax: 41446355712
Nazionalità Coordinatore	Switzerland [CH]
Totale costo	184˙709 €
EC contributo	184˙709 €
Programma	FP7-PEOPLE Specific programme "People" implementing the Seventh Framework Programme of the European Community for research, technological development and demonstration activities (2007 to 2013)
Code Call	FP7-PEOPLE-2011-IEF
Funding Scheme	MC-IEF
Anno di inizio	2012
Periodo (anno-mese-giorno)	2012-12-01   -   2014-11-30

 Partecipanti

#	participant	country	role	EC contrib. [€]
1	UNIVERSITAET ZUERICH  Organization address address: Raemistrasse 71 city: ZURICH postcode: 8006 contact info Titolo: Prof. Nome: Andreas Cognome: Plückthun Email: send email Telefono: 41446355571 Fax: 41446355712	CH (ZURICH)	coordinator	184˙709.40

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 Obiettivo del progetto (Objective)

'Class B G-protein coupled receptors (GPCRs) are peptide hormone-binding receptors that are implicated in the pathogenesis several human diseases which makes them attractive targets for drug therapy. To develop new compounds to target these receptors a detailed understanding of the molecular structure is required which has not been succeeded to date. To elucidate the structure of proteins by X-ray crystallography or NMR spectroscopy large quantities of protein are required. For GPCRs this prerequisite is difficult to achieve as the vast majority of GPCRs exhibits low endogenous expression and is very unstable in solution. Therefore, improved expression conditions are required for the efficient characterization of new GPCR structures. In the proposed project class B GPCRs will be optimized for improved heterologous expression and increased thermostability by means of directed evolution. Libraries of class B GPCRs will be obtained by random mutagenesis and will be heterologously expressed using a system in which functional GPCR is targeted to the inner membrane of E. coli. Mutants that display increased receptor expression levels and ligand binding will be selected by flow cytometry using fluorescently labeled ligands. Repetitive cycles of randomization and selection will allow to gradually increasing the level of protein expression and stability. With this evolutionary approach key residues within the receptor sequence can be rapidly identified that are responsible for improved biophysical properties without affecting the pharmacological properties of the receptor. Such GPCR mutants will be a valuable tool on the way to express high quantities of stable receptor protein for subsequent structural studies.'

Introduzione (Teaser)

To develop a new drug one needs to know the detailed molecular structure of a targeted protein. An EU study aimed to optimise heterologous protein expression to obtain a more stable protein needed for a structural study.