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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)

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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.

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Abbreviations

DR5:

Death receptor 5

TRAIL:

Tumor necrosis factor-a-Related Apoptosis-Inducing Ligand

ECD:

Extra cellular domain

HSQC:

Heteronuclear single quantum coherence transfer

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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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Authors and Affiliations

  1. Chimie et Biologie des Membranes et des Nano-objets (CBMN), Université de Bordeaux – CNRS – Bordeaux INP, UMR 5248, Bâtiment B14, Allée Geoffroy Saint Hilaire, 33600, Pessac Cedex, France

    Antoine Baudin, Anne Guichard, Gavin W. Collie, Stéphane Chaignepain, Agnès Hocquellet, Mélanie Berbon, Antoine Loquet, Gilles Guichard & Benoît Odaert

  2. Agenus UK Limited, 315, Science Park, Milton Road, Cambridge, CB4 0WG, UK

    Anne Guichard

  3. Institut Européen de Chimie et Biologie, Univ. Bordeaux, 2 rue Robert Escarpit, 33607, Pessac, France

    Gavin W. Collie, Sabrina Rousseau, Mélanie Berbon, Antoine Loquet, Cameron Mackereth & Gilles Guichard

  4. Centre de Génomique Fonctionnelle de Bordeaux (CGFB), 146 rue Léo Saignat, 33000, Bordeaux, France

    Stéphane Chaignepain

  5. Inserm U1212, CNRS UMR5320, ARNA Laboratory, 146 rue Léo Saignat, 33076, Bordeaux, France

    Sabrina Rousseau & Cameron Mackereth

  6. Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, UK

    Gavin W. Collie

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  1. Antoine Baudin
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  2. Anne Guichard
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  3. Gavin W. Collie
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  8. Antoine Loquet
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  10. Gilles Guichard
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  11. Benoît Odaert
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Correspondence to Benoît Odaert.

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Baudin, A., Guichard, A., Collie, G.W. et al. 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). Biomol NMR Assign 12, 309–314 (2018). https://doi.org/10.1007/s12104-018-9828-1

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  • DOI: https://doi.org/10.1007/s12104-018-9828-1

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