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Nanobody-enabled Structural Investigation of a G Protein-Coupled Receptor-Arrestin Complex

Objective

Although many thousands of transient protein-protein interactions (PPIs) are known, there is a disturbing paucity of high-resolution structures of the resulting complexes and the difficulties involved in experimentally determining these atomic structures remain essentially unaddressed.

The Steyaert lab has shown that cross-linking transiently interacting proteins, followed by immunization of llama’s with this cross-linked antigen, causes the maturation of single domain antibodies called Nanobodies (Nbs). The Nbs bind composite conformational epitopes unique to the transient complex. Highly efficient selection methods can discriminate Nbs that exclusively bind the transient (non-cross-linked) complex from binders that bind to the dissociated monomers. Such Nbs will be instrumental to purify and solve the structures of PPIs that have been resistant to investigation by X-ray, NMR, SAXS or EM and for the functional analysis of these complexes within a living cell.

This ground-breaking technology will be validated with well-chosen case studies covering key PPIs of the GPCR transmembrane signaling pathway including parts of the arrestin interactome. During this project the fellow will establish a unique research niche by systematically applying her arsenal of learnt techniques to develop the next generation antibody‐enabled methods for the structural investigation of the GPCR-arrestin targets. The ultimate goal of NESIAC is to determine the atomic structures of the most relevant transient associations of these signaling proteins. This will open up a new platform for realising the structural basis of the elusive GPCR regulation by arrestins.

Fields of science (EuroSciVoc)

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Programme(s)

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Topic(s)

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Funding Scheme

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MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)

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Call for proposal

Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.

(opens in new window) H2020-MSCA-IF-2014

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Coordinator

VIB VZW
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 160 800,00
Address
SUZANNE TASSIERSTRAAT 1
9052 ZWIJNAARDE - GENT
Belgium

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Region
Vlaams Gewest Prov. Oost-Vlaanderen Arr. Gent
Activity type
Research Organisations
Links
Total cost

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.

€ 160 800,00
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