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New Routes for the Solution NMR Investigations of Extra Large Biomolecular Assemblies

Objective

The cell is a collection of dynamic molecular machines. NMR spectroscopy is the method of choice to observe, at atomic resolution, complex conformational changes, transient interactions and dynamics of proteins. Introduction of methyl specific labelling technology has enabled solution NMR studies of protein assemblies of several hundred kDa. However, this strategy is mostly restricted to symmetrical and thermostable protein assemblies, precluding applications to large medically relevant biological complexes. The increases of linewidths and the high number of signal overlaps hamper the NMR spectra analysis of most hetero-oligomeric large assemblies studied at room temperature. In this project, we will develop two complementary concepts to significantly simplify both NMR spectra and corresponding site specific analysis. We will build a multi-site specific labelling method enabling observation of NMR signals only for the sites of interest in a large protein complex. We will invent a combinatorial strategy to reduce the time required for site-specific identification of each individual NMR signal from a few months to a few hours. New 1H-frequencies edition schemes will be introduced to enhance significantly the NMR spectras resolution of very slow tumbling biological particles. These groundbreaking NMR methods will be validated using therapeutic antibodies (150 kDa), ribosome (2.4 MDa) samples, and will be used directly to capture the mechanisms of ATP-fueled human chaperonin (1 MDa) in complex with the aggregation prone form of Huntingtin. This project will provide new technological breakthroughs to push biological applications of NMR significantly beyond its current boundaries. We anticipate that the simplification of the NMR analysis resulting from the new developed NMR routes will transform the solution NMR spectroscopy in a very competitive method to study large medically relevant biomolecular assemblies and molecular machines, so far considered as untargetable.

Fields of science (EuroSciVoc)

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Keywords

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Programme(s)

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Topic(s)

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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.

HORIZON-ERC - HORIZON ERC Grants

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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) ERC-2022-ADG

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Host institution

CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE CNRS
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.

€ 2 500 000,00
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.

€ 2 500 000,00

Beneficiaries (2)

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