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Biomolecular NMR Assignments

, Volume 12, Issue 2, pp 309–314 | Cite as

1H, 13C, 15N NMR resonance assignments and secondary structure determination of the extra-cellular domain from the human proapoptotic TRAIL-R2 death receptor 5 (DR5-ECD)

  • Antoine Baudin
  • Anne Guichard
  • Gavin W. Collie
  • Sabrina Rousseau
  • Stéphane Chaignepain
  • Agnès Hocquellet
  • Mélanie Berbon
  • Antoine Loquet
  • Cameron Mackereth
  • Gilles Guichard
  • Benoît OdaertEmail author
Article
  • 165 Downloads

Abstract

Death receptors (DR) selectively drive cancer cells to apoptosis upon binding to the Tumor necrosis factor-a-Related Apoptosis-Inducing Ligand (TRAIL). Complex formation induces the oligomerization of the death receptors DR4 (TRAIL-R1) and DR5 (TRAIL-R2) and transduces the apoptogenic signal to their respective death domains, leading to Death Inducing Signaling Complex (DISC) formation, caspase activation and ultimately cell death. Several crystal structures of the ExtraCellular Domain from Death Receptor 5 (DR5-ECD) have been reported in complex with the TRAIL ligand or anti-DR5 antibodies, but none for the isolated protein. In order to fill this gap and to perform binding experiments with TRAIL peptidomimetics, we have produced isotopically labelled DR5-ECD and started a conformational analysis by using high-field 3D NMR spectroscopy. Herein, we present the first resonance assignment of a TRAIL receptor in solution and the determination of its secondary structure from NMR chemical shifts.

Keywords

DR5 TRAIL Cancer Apoptosis NMR spectroscopy Resonance assignment Secondary structure 

Abbreviations

DR5

Death receptor 5

TRAIL

Tumor necrosis factor-a-Related Apoptosis-Inducing Ligand

ECD

Extra cellular domain

HSQC

Heteronuclear single quantum coherence transfer

Notes

Acknowledgements

We thank the structural biology platform at the Institut Européen de Chimie et Biologie (UMS 3033) for access to NMR spectrometers and technical assistance. Antoine Baudin was supported by a French PhD fellowship afforded to the University of Bordeaux by the Ministère de la Recherche (MNERT) and a fourth year PhD extension by the Ligue nationale Contre le Cancer (LCC). The project was funded by the Ligue Contre le Cancer de la Gironde. We thank Dr Yong-Sung Kim from the Ajou University of the Republic of Korea for providing us the plasmid containing the gene coding for DR5-ECD. AL and MB acknowledge the H2020 program (ERC-2015-StG GA no. 639020).

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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Antoine Baudin
    • 1
  • Anne Guichard
    • 1
    • 2
  • Gavin W. Collie
    • 1
    • 3
    • 6
  • Sabrina Rousseau
    • 3
    • 5
  • Stéphane Chaignepain
    • 1
    • 4
  • Agnès Hocquellet
    • 1
  • Mélanie Berbon
    • 1
    • 3
  • Antoine Loquet
    • 1
    • 3
  • Cameron Mackereth
    • 3
    • 5
  • Gilles Guichard
    • 1
    • 3
  • Benoît Odaert
    • 1
    Email author
  1. 1.Chimie et Biologie des Membranes et des Nano-objets (CBMN)Université de Bordeaux – CNRS – Bordeaux INP, UMR 5248Pessac CedexFrance
  2. 2.Agenus UK LimitedCambridgeUK
  3. 3.Institut Européen de Chimie et BiologieUniv. BordeauxPessacFrance
  4. 4.Centre de Génomique Fonctionnelle de Bordeaux (CGFB)BordeauxFrance
  5. 5.Inserm U1212, CNRS UMR5320, ARNA LaboratoryBordeauxFrance
  6. 6.Discovery Sciences, IMED Biotech UnitAstraZenecaCambridgeUK

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