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Project

Nanobody-enabled structural and functional studies of Chemokine•CCR5 active-state complexes (FWOAL804)

G protein-coupled receptors (GPCRs) are highly dynamic membrane receptors existing in many conformational states. Binding of agonists at the GPCR extracellular side stabilizes their active state triggering intracellular signaling. Among the largest GPCR ligands are the chemokines, most notably known for their role in guiding cell migration during development and immune responses. The chemokine-receptor interaction network is highly complex and often chemokines with little sequence identity bind and activate the same receptor. CCR5, which binds three chemokines as well as the HIV envelope proteins, is an example of such a receptor. The determinants regulating CCR5 ligand selectivity and activation are unclear.

Structural investigation of chemokine receptors bound to their bulky agonists is difficult due to their inherent instability. We have previously shown the remarkable capacity of Nanobodies, the antigen-binding fragments of heavy-chain-only antibodies from camelids, to stabilize GPCR active states or GPCR-protein assemblies.

In this project, by applying an original approach of presenting CCR5 bound to its different ligands on artificial membranes (liposomes) we aim at generating Nanobodies stabilizing the receptor active states to investigate the crosstalk between the extracellular ligand-binding site and the intracellular signaling protein-binding site and ultimately solving the structure of CCR5-ligand complexes by Nanobody-assisted X-ray crystallography.
Date:1 Jan 2016 →  31 Dec 2019
Keywords:GPCR
Disciplines:General chemical and biochemical engineering not elsewhere classified