Project description
Improved protein labelling supports studies of dynamics at distances
Proteins perform a plethora of functions in cells and tissues, many accomplished via conformational changes. Conversely, aberrant changes in protein structure are often linked to pathological conditions. Paramagnetic resonance enhancements (PRE) or single-molecule Förster resonance energy transfer (smFRET), two advanced spectroscopic techniques for studying real-time protein dynamics in solutions relying on site-specific labelling, have enabled measurements of interactions over different distances. Current labelling approaches dominantly rely on random cysteine labelling with only a handful of probes. With the support of the Marie Skłodowska-Curie Actions programme, the InProSpecT project aims to develop tailored, selective and easy-to-use fluorophores and labelling strategies for smFRET and PRE, allowing simultaneous studies of protein distances and dynamics.
Objective
Protein structures, including their complexes, determine their specific function and the molecular working mechanisms. The astonishing developments in X-ray crystallography and cryo-EM allow insights into atomic levels of protein structures, however, commonly on their static, solid state conformations. Nuclear magnetic resonance (NMR) and single molecule fluorescence spectroscopy, on the other hand, allow observations of protein dynamics in solution, in real-time. In particular, specific intermediate and long-range interactions can be measured using paramagnetic resonance enhancements (PRE) or single molecule Frster
Resonance Energy Transfer (smFRET), requiring site-specific labelling with paramagnetic or fluorescence probes, respectively.
Naturally, the choice of the label and its covalent attachment to the protein are critical, however, traditional approaches rely on random cysteine labelling with only a handful of available probes, making measurements difficult to interpret and laborious. As such, there is an urgent demand to develop custom-tailored, selective, orthogonal, easy to use chemical biology probes with excellent spectroscopic properties (for smFRET and PRE) to drive the field of protein structure and dynamics. This proposal tackles current limitations in the peptide/protein labelling field by creating state-of-the-art fluorophores for red/far-red smFRET. Building on this, the best performing dyes will be unleashed to defined labelling of intrinsically disordered and native proteins, in vitro and in cellulo, respectively. Next, these concepts will be expanded to be merged with intramolecular PRE probes to combine to date mutually exclusive spectroscopic properties. InProSpecT (Innovative Protein labelling strategies for Spectroscopic high resolution Techniques) pushes a bottom-up approach, from fluorophore design and labelling strategies ultimately linking two worlds to allow unprecedented, simultaneous studies of protein distances and dynamics.
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.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques.
- natural sciencesearth and related environmental sciencesgeologymineralogycrystallography
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteins
- natural sciencesphysical sciencesopticsspectroscopy
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Keywords
Programme(s)
- HORIZON.1.2 - Marie Skłodowska-Curie Actions (MSCA) Main Programme
Funding Scheme
HORIZON-TMA-MSCA-PF-EF - HORIZON TMA MSCA Postdoctoral Fellowships - European FellowshipsCoordinator
12489 Berlin
Germany