Abstract
Nanobodies are single chain antibodies that have become a highly valuable and versatile tool for biomolecular and therapeutic research. One application field is the stabilization of active states of flexible proteins, among which G-protein coupled receptors represent a very important class of membrane proteins. Here we present the backbone and side-chain assignment of the 1H, 13C and 15N resonances of Nb33 and Nb39, two nanobodies that recognize and stabilize the µ-opioid receptor to opioids in its active agonist-bound conformation. In addition, we present a comparison of their secondary structures as derived from NMR chemical shifts.
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De Genst E et al (2006) Molecular basis for the preferential cleft recognition by dromedary heavy-chain antibodies. Proc Natl Acad Sci U S A 103:4586–4591. doi:10.1073/pnas.0505379103
Delaglio F, Grzesiek S, Vuister GW, Zhu G, Pfeifer J, Bax A (1995) NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR 6:277–293
Deupi X, Kobilka BK (2010) Energy landscapes as a tool to integrate GPCR structure, dynamics, and function. Physiology 25:293–303. doi:10.1152/physiol.00002.2010
Hamers-Casterman C et al (1993) Naturally-occurring antibodies devoid of light-chains. Nature 363:446–448. doi:10.1038/363446a0
Huang W et al (2015) Structural insights into micro-opioid receptor activation. Nature 524:315–321. doi:10.1038/nature14886
Markley JL et al (1998) Recommendations for the presentation of NMR structures of proteins and nucleic acids–IUPAC-IUBMB-IUPAB Inter-Union Task Group on the standardization of data bases of protein and nucleic acid structures determined by NMR spectroscopy. Eur J Biochem 256:1–15
Melnikova I (2010) Pain market. Nat Rev Drug Discov 9:589–590. doi:10.1038/nrd3226
Neilan CL et al (2004) Characterization of the complex morphinan derivative BU72 as a high efficacy, long-lasting mu-opioid receptor agonist. Eur J Pharmacol 499:107–116. doi:10.1016/j.ejphar.2004.07.097
Pons JL, Malliavin TE, Delsuc MA (1996) Gifa V. 4: a complete package for NMR data set processing. J Biomol NMR 8:445–452. doi:10.1007/BF00228146
Salema V, Fernandez LA (2013) High yield purification of nanobodies from the periplasm of E. coli as fusions with the maltose binding protein. Protein Expr Purif 91:42–48. doi:10.1016/j.pep.2013.07.001
Shen Y, Bax A (2013) Protein backbone and sidechain torsion angles predicted from NMR chemical shifts using artificial neural networks. J Biomol NMR 56:227–241. doi:10.1007/s10858-013-9741-y
Sounier R et al (2015) Propagation of conformational changes during mu-opioid receptor activation. Nature 524:375–378. doi:10.1038/nature14680
Van Audenhove I, Gettemans J (2016) Nanobodies as versatile tools to understand, diagnose, visualize and treat cancer. EBioMedicine 8:40–48. doi:10.1016/j.ebiom.2016.04.028
Vranken WF et al (2005) The CCPN data model for NMR spectroscopy: development of a software pipeline. Proteins 59:687–696. doi:10.1002/prot.20449
Wishart DS et al (1995) 1 H, 13 C and 15 N chemical shift referencing in biomolecular NMR. J Biomol NMR 6:135–140
Acknowledgements
This work used the NMR FRISBI (French Infrastructure for Integrated Structural Biology) platform in Montpellier, with support from the “Agence nationale de la recherche” of France, ANR-10-INSB-05-0 (YY and HD). We acknowledge the GIS “IBiSA: Infrastructures en Biologie Santé et Agronomie” and the support from the National Institutes of Health Grant (NIDA-DA036246 to SG).
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Sounier, R., Yang, Y., Hagelberger, J. et al. 1H, 13C and 15N backbone chemical shift assignments of camelid single-domain antibodies against active state µ-opioid receptor. Biomol NMR Assign 11, 117–121 (2017). https://doi.org/10.1007/s12104-017-9733-z
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DOI: https://doi.org/10.1007/s12104-017-9733-z