Preprints
https://doi.org/10.5194/mr-2021-54
https://doi.org/10.5194/mr-2021-54

  12 Jul 2021

12 Jul 2021

Review status: a revised version of this preprint was accepted for the journal MR and is expected to appear here in due course.

Fluorine NMR study of proline-rich sequences using fluoroprolines

Davy Sinnaeve1,2, Abir Ben Bouzayene3, Emile Ottoy4, Gert-Jan Hofman4,5, Eva Erdmann3, Bruno Linclau5, Ilya Kuprov5, José C. Martins4, Vladimir Torbeev6, and Bruno Kieffer3 Davy Sinnaeve et al.
  • 1Univ. Lille, Inserm, Institut Pasteur de Lille, CHU Lille, U1167 – Labex DISTALZ – RID-AGE – Risk Factors and Molecular Determinants of Aging-Related Diseases, F-59000 Lille, France
  • 2CNRS, ERL9002 - Integrative Structural Biology, F-59000 Lille, France
  • 3Departement of Integrative Structural Biology, IGBMC, Université de Strasbourg, INSERM U1258, CNRS UMR7104, 1, rue Laurent Fries, F-67404 Illkirch, France
  • 4Department of Organic and Macromolecular Chemistry, Ghent University, Campus Sterre, S4, Krijgslaan 281, B-9000 Gent, Belgium
  • 5School of Chemistry, University of Southampton, Southampton SO17 1BJ, United Kingdom
  • 6Institut de Science et d’Ingénierie Supramoléculaires (ISIS), International Center for Frontier Research in Chemistry (icFRC), University of Strasbourg, CNRS UMR 7006, F-67000 Strasbourg, France

Abstract. Proline homopolymer motifs are found in many proteins; their peculiar conformational and dynamic properties are often directly involved in those proteins' functions. However, the dynamics of proline homopolymers is hard to study by NMR due to lack of amide protons and small chemical shift dispersion. Exploiting the spectroscopic properties of fluorinated prolines opens interesting perspectives to address these issues. Fluorinated prolines are already widely used in protein structure engineering – they introduce conformational and dynamical biases – but their use as 19F NMR reporters of proline conformation has not yet been explored. In this work, we look at model peptides where Cγ-fluorinated prolines with opposite configurations of the chiral Cγ centre have been introduced at two postions in distinct polyproline segments. By looking at the effects of swapping these (4R)- and (4S)-4-fluoroprolines witin the polyproline segments, we were able to separate the intrinsic conformational properties of the polyproline sequence from the conformational alterations instilled by fluorination. We assess the fluoroproline 19F relaxation properties, and exploit the latter in elucidating binding kinetics to the SH3 domain.

Davy Sinnaeve et al.

Status: closed

Comment types: AC – author | RC – referee | CC – community | EC – editor | CEC – chief editor | : Report abuse
  • RC1: 'Comment on mr-2021-54', Ranajeet Ghose, 19 Jul 2021
    • AC1: 'Reply on RC1', Bruno Kieffer, 09 Sep 2021
  • RC2: 'Comment on mr-2021-54', Guy Lippens, 30 Jul 2021
    • AC2: 'Reply on RC2', Bruno Kieffer, 09 Sep 2021
  • RC3: 'Comment on mr-2021-54', Anonymous Referee #3, 10 Aug 2021
    • AC3: 'Reply on RC3', Bruno Kieffer, 09 Sep 2021

Status: closed

Comment types: AC – author | RC – referee | CC – community | EC – editor | CEC – chief editor | : Report abuse
  • RC1: 'Comment on mr-2021-54', Ranajeet Ghose, 19 Jul 2021
    • AC1: 'Reply on RC1', Bruno Kieffer, 09 Sep 2021
  • RC2: 'Comment on mr-2021-54', Guy Lippens, 30 Jul 2021
    • AC2: 'Reply on RC2', Bruno Kieffer, 09 Sep 2021
  • RC3: 'Comment on mr-2021-54', Anonymous Referee #3, 10 Aug 2021
    • AC3: 'Reply on RC3', Bruno Kieffer, 09 Sep 2021

Davy Sinnaeve et al.

Davy Sinnaeve et al.

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Short summary
Fluorine NMR was used to study the interaction between a proline-rich peptide and a SH3 domain using 4S- and 4R fluorinated prolines whose potential as NMR probes has not been exploited yet. We present a comprehensive study addressing several aspects to be considered when using these residues as NMR probes, including relaxation and dynamics. We show that their conformational bias may be used to modulate the kinetics of protein binding to proline-rich motifs.