Objective
G protein-coupled receptors (GPCRs) are eukaryotic seven-alpha-helix transmembrane proteins which are involved in extracellular signal transduction across the cellular membrane. Aim of the proposed research is to structurally characterise ligand-induced conformational changes in GPCRs, to identify the number of alternative conformations the GPCR can adopt, and to engineer conformationally stabilised GPCR variants for subsequent biophysical and crystallographic studies. The research will focus on cannabinoid receptor type 2 and beta-1 adrenergic receptor. Both GPCRs can be expressed and purified from recombinant Escherichia coli as well as insect and mammalian cells. Degree of ligand-induce conformational change will be determined for each amino acid residue by using a novel protein-engineering approach that combines alanine scanning mutagenesis and conformational phi-value analysis. Transition temperatures of denaturation for native and each mutated protein will be determined in presence and absence of particular ligand by high-throughput differential scanning fluorimetry or radioligand-binding assays. These results will be used in engineering of GPCR mutants that are more stable and locked in the specific (eg, activated or inactive) conformational state. Ligand-binding to the stabilised mutants and their functionality will be monitored by fluorescence anisotropy titrations, as well as by radioactive assays using the corresponding G proteins. The stable and functional variants will be used in crystallisation trials. The diffraction data from the expected microcrystals will be collected on the microfocus X-ray beamline. Crystal structures will be determined by molecular replacement. The proposed research will give insights into the activation mechanisms of GPCRs. Detailed understanding of the conformational changes accompanying receptor activation is a prerequisite for rational design of therapeutic drugs affecting GPCRs.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- engineering and technology materials engineering crystals
- natural sciences biological sciences biochemistry biomolecules proteins
- natural sciences biological sciences zoology entomology
- natural sciences chemical sciences organic chemistry amines
You need to log in or register to use this function
Programme(s)
Multi-annual funding programmes that define the EU’s priorities for research and innovation.
Multi-annual funding programmes that define the EU’s priorities for research and innovation.
Topic(s)
Calls for proposals are divided into topics. A topic defines a specific subject or area for which applicants can submit proposals. The description of a topic comprises its specific scope and the expected impact of the funded project.
Calls for proposals are divided into topics. A topic defines a specific subject or area for which applicants can submit proposals. The description of a topic comprises its specific scope and the expected impact of the funded project.
Call for proposal
Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.
Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.
FP7-PEOPLE-2010-IEF
See other projects for this call
Funding Scheme
Funding scheme (or “Type of Action”) inside a programme with common features. It specifies: the scope of what is funded; the reimbursement rate; specific evaluation criteria to qualify for funding; and the use of simplified forms of costs like lump sums.
Funding scheme (or “Type of Action”) inside a programme with common features. It specifies: the scope of what is funded; the reimbursement rate; specific evaluation criteria to qualify for funding; and the use of simplified forms of costs like lump sums.
Coordinator
5232 VILLIGEN PSI
Switzerland
The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.