Titre :

A library of fluorinated prolines to study proline−rich motifs with 19F NMR

Auteur(s) :

Sinnaeve, Davy [Orateur]
Facteurs de Risque et Déterminants Moléculaires des Maladies liées au Vieillissement - U 1167 [RID-AGE]
Biologie Structurale Intégrative [ERL 9002 - INSERM U1167 - BSI]
Ottoy, Emile [Auteur]
Linclau, Bruno [Auteur]
Kieffer, Bruno [Auteur]
Institut de Génétique et de Biologie Moléculaire et Cellulaire [IGBMC]
Kuprov, Ilya [Auteur]
School of Chemistry [Southampton, UK]

Titre de la manifestation scientifique :

Advanced Isotopic Labelling Methods for Integrated Structural Biology 2024

Ville :

Grenoble

Pays :

France

Date de début de la manifestation scientifique :

2024-05-28

Date de publication :

2024-05-31

Mot(s)-clé(s) en anglais :

NMR spectroscopy
Fluorine NMR

Discipline(s) HAL :

Chimie/Chimie analytique
Chimie/Chimie organique
Chimie/Chimie théorique et/ou physique
Sciences du Vivant [q-bio]/Biochimie, Biologie Moléculaire/Biologie structurale [q-bio.BM]

Résumé en anglais : [en]

Proline-rich motifs (PRMs) often occur in intrinsically disordered proteins, often forming sites for protein-protein interactions and thus heavily involved in organizing protein structures and key to many cell signaling ...
Lire la suite >Proline-rich motifs (PRMs) often occur in intrinsically disordered proteins, often forming sites for protein-protein interactions and thus heavily involved in organizing protein structures and key to many cell signaling processes and diseases. They are often challenging to study with NMR due to the lack of an amide proton and the limited signal dispersion of its aliphatic side-chain, especially in low-complexity sequences or in oligoproline stretches. We propose fluorinated prolines (FPros)[1] as an attractive tool to question the role of individual proline residues. Firstly, 19F NMR provides easy spectroscopic access to specific residues, revealing for instance the cis-trans ratio.[2] Secondly, fluorination alters proline’s conformational dynamics, including the five-membered ring conformation of the side chain and the cis-trans ratio. This occurs in a way that depends on the precise stereospecific fluorine substitution. This form of conformational pre-organization has previously been very instructive to reveal the link between proline conformation and overall protein stability in folded proteins,[1] with collagen as a landmark example.[3] To make full use of both the 19F NMR and pre-organizing abilities of FPros, we have characterized all 10 mono- and difluoroprolines substituted at the 3- and/or 4-positions using NMR on synthetic model compounds,[4,5] supplemented with an extensive full conformational landscape analysis of their five-membered rings using Density Functional Theory. This allows full mapping of the 1H and 19F NMR properties as a function of conformation, such as 19F CSA and J-couplings. We will present how these FPro residues can be put to good use as 19F NMR reporters in PRMs on a number of examples.REFERENCES:1.Verhoork S.J.M., Killoran P.M. and Coxon C.R. Fluorinated Prolines as Conformational Tools and Reporters for Peptide and Protein Chemistry Biochemistry, 57, 6132-6143 (2018).2.Sinnaeve D., Ben Bouzayene A., Ottoy E., Hofman G.J., Erdmann E., Linclau B., Kuprov I., Martins J.C., Torbeev V. and Kieffer B. Fluorine NMR study of proline-rich sequences using fluoroprolines Magnetic Resonance, 2, 795-813 (2021).3.Bretscher L.E., Jenkins C.L., Taylor K.M., DeRider M.L. and Raines R.T. Conformational stability of collagen relies on a stereoelectronic effect Journal of the American Chemical Society, 123, 777-778 (2001).4.Hofman G.J., Ottoy E., Light M.E., Kieffer B., Kuprov I., Martins J.C., Sinnaeve D. and Linclau B. Minimising conformational bias in fluoroprolines through vicinal difluorination Chemical Communications, 54, 5118-5121 (2018).5.Hofman G.J., Ottoy E., Light M.E., Kieffer B., Kuprov I., Martins J.C., Sinnaeve D. and Linclau B. Synthesis and Conformational Properties of 3,4-Difluoro-L-prolines Journal of Organic Chemistry, 84, 3100-3120 (2019).Lire moins >

Langue :

Anglais

Comité de lecture :

Oui

Audience :

Internationale

Vulgarisation :

Non

Collections :

• Facteurs de risque et déterminants moléculaires des maladies liées au vieillissement (RID-AGE) - U1167

Source :

Harvested from HAL

Date de dépôt :

2025-04-03T03:51:41Z