Harvesting the potential of fluorine labels and NMR spectroscopy to study the conformation of polyproline sequences and their interaction with biomolecular targets

01 January 2015 → 31 December 2020
Research Foundation - Flanders (FWO)
Research disciplines
  • Natural sciences
    • Organic chemistry
NMR spectroscopy polyproline sequences biomolecular targets fluorine labels
Project description

Proline-rich or polyproline regions play an important role in biology, often involved in proteinprotein interactions that mediate cell signaling or gene regulation. Typically present in disordered protein domains, their characterization by NMR spectroscopy is often hampered by the lack of an amide proton in Pro residues and the low resolution between resonances of the different residues. The otherwise so powerful ability of NMR to probe residue specific information is therefore lost. We propose to introduce fluorinated proline (FPro) residues, making use of both the 19F spin label’s strong sensitivity to molecular environment and the significant perturbation it creates on 1H and 13C chemical shifts within the residue. By considering various possible mono- and difluorinaed FPros we wish to establish a strategic approach that recovers NMR as a tool to study the conformational and dynamical properties of proline-rich peptide sequences. The investigation involves both the organic synthesis of novel fluorinated proline residues and the assessment of the conformational and dynamical behavior relative to native proline. Next to the methodological development, our approach will be directly applied on a biochemical case study. Here, the impact of Ser phosphorylation on the conformational landscape of a proline-rich modelpeptide mimicking the retinoic acid nuclear receptor will be investigated, a study relevant to understand hormone mediated gene regulation.