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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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.
KeywordsDR5 TRAIL Cancer Apoptosis NMR spectroscopy Resonance assignment Secondary structure
Death receptor 5
Tumor necrosis factor-a-Related Apoptosis-Inducing Ligand
Extra cellular domain
Heteronuclear single quantum coherence transfer
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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