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Pinpoint-NMR: A sequence-specific labeling technique to accelerate NMR investigation of large proteins

Project description

Advanced NMR spectroscopy for studying large protein complexes

Nuclear magnetic resonance (NMR) spectroscopy is an analytical technique that reveals the atoms and how they interact within molecules. Researchers rely on NMR to examine protein structure and understand their function in health and disease. However, studying large protein complexes remains challenging due to signal overlap and intricate sample preparation. With the support of the Marie Skłodowska-Curie Actions programme, the PINPOINT-NMR project introduces an innovative method for labelling specific sites within large proteins. This targeted approach promises to simplify NMR spectra and accelerate analysis. As a result, it will make it easier to study large biomolecules such as therapeutic antibodies, supporting biomedical research and drug development.

Objective

The cell is an ensemble of dynamic molecular machines. NMR spectroscopy is the method of choice to observe complex conformational changes, transient interactions, and dynamics of proteins at atomic resolution. The introduction of methyl-specific labeling has made it possible to study protein assemblies of several hundred kDa by solution NMR. However, the high number of overlapping signals and the labor-intensive process of site specific analysis hamper NMR investigation of large hetero-oligomeric assemblies at room temperature. While segmental labeling or site-specific labeling offer opportunities to simplify 2D NMR spectra, the complexity of sample preparation or the characteristic low expression yield drastically limit its subsequent applications. Therefore, efficient, widely applicable, and straightforward labeling strategies are necessary to integrate solution NMR analysis of large proteins (such as monoclonal antibodies) into R&D pipelines for biomedical industry. I propose to harness the natural (anti)codon code for multi-site, sequence-specific isotope labeling. The resulting method would simplify the spectral congestion typical of 2D NMR spectra of large proteins, allowing targeted investigation of sites of interest (e.g. catalytic centers) and streamlining residue assignment. This method will reduce the time required for residue specific analysis of NMR data from months to days, thus expanding our ability to study large, medically-relevant complexes and address current biological challenges.

Fields of science (EuroSciVoc)

Keywords

Programme(s)

Topic(s)

Funding Scheme

HORIZON-TMA-MSCA-PF-EF - HORIZON TMA MSCA Postdoctoral Fellowships - European Fellowships

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Call for proposal

(opens in new window) HORIZON-MSCA-2024-PF-01

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Coordinator

UNIVERSITE GRENOBLE ALPES
Net EU contribution
€ 283 025,70
Address
621 AVENUE CENTRALE
38058 Grenoble
France
Region
Auvergne-Rhône-Alpes Rhône-Alpes Isère
Activity type
Higher or Secondary Education Establishments
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Total cost
No data

Partners (2)