6VI4

Nanobody-Enabled Monitoring of Kappa Opioid Receptor States

Summary for 6VI4

• Entry DOI: 10.2210/pdb6vi4/pdb
• Descriptor: Kappa opioid receptor, Nanobody 6, (3R)-7-hydroxy-N-{(2S)-1-[(3R,4R)-4-(3-hydroxyphenyl)-3,4-dimethylpiperidin-1-yl]-3-methylbutan-2-yl}-1,2,3,4-tetrahydroisoquinoline-3-carboxamide, ... (4 entities in total)
• Functional Keywords: g protein-coupled opioid receptor, membrane protein
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 4
• Total formula weight: 100409.22

Authors

Che, T.,Roth, B.L. (deposition date: 2020-01-11, release date: 2020-03-18, Last modification date: 2024-11-13)

Primary citation

Che, T.,English, J.,Krumm, B.E.,Kim, K.,Pardon, E.,Olsen, R.H.J.,Wang, S.,Zhang, S.,Diberto, J.F.,Sciaky, N.,Carroll, F.I.,Steyaert, J.,Wacker, D.,Roth, B.L.
Nanobody-enabled monitoring of kappa opioid receptor states.
Nat Commun, 11:1145-1145, 2020

PubMed Abstract: Recent studies show that GPCRs rapidly interconvert between multiple states although our ability to interrogate, monitor and visualize them is limited by a relative lack of suitable tools. We previously reported two nanobodies (Nb39 and Nb6) that stabilize distinct ligand- and efficacy-delimited conformations of the kappa opioid receptor. Here, we demonstrate via X-ray crystallography a nanobody-targeted allosteric binding site by which Nb6 stabilizes a ligand-dependent inactive state. As Nb39 stabilizes an active-like state, we show how these two state-dependent nanobodies can provide real-time reporting of ligand stabilized states in cells in situ. Significantly, we demonstrate that chimeric GPCRs can be created with engineered nanobody binding sites to report ligand-stabilized states. Our results provide both insights regarding potential mechanisms for allosterically modulating KOR with nanobodies and a tool for reporting the real-time, in situ dynamic range of GPCR activity.

PubMed: 32123179
DOI: 10.1038/s41467-020-14889-7

Experimental method

X-RAY DIFFRACTION (3.3 Å)