PDBsum entry 6vi4

Membrane protein

PDB id: 6vi4

Contents

Protein chains

283 a.a.

118 a.a.

Ligands

JDC ×2

CLR

PDB id:

6vi4

Name:

Membrane protein

Title:

Nanobody-enabled monitoring of kappa opioid receptor states

Structure:

Kappa opioid receptor. Chain: a, b. Synonym: kor-1. Engineered: yes. Mutation: yes. Nanobody 6. Chain: c, d. Engineered: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: oprk1, oprk. Expressed in: spodoptera frugiperda. Expression_system_taxid: 7108. Lama glama. Organism_taxid: 9844. Expressed in: escherichia coli.

Resolution:

3.30Å R-factor: 0.243 R-free: 0.271

Authors:

T.Che,B.L.Roth

Key ref:

T.Che et al. (2020). Nanobody-enabled monitoring of kappa opioid receptor states. Nat Commun, 11, 1145. PubMed id: 32123179 DOI: 10.1038/s41467-020-14889-7

Date:

11-Jan-20 Release date: 18-Mar-20

Protein chains

P41145  (OPRK_HUMAN) -  Kappa-type opioid receptor from Homo sapiens

Seq: 380 a.a.

Struc: 283 a.a.*

Protein chains

No UniProt id for this chain

Struc: 118 a.a.

* PDB and UniProt seqs differ at 1 residue position (black cross)

DOI no: 10.1038/s41467-020-14889-7

Nat Commun 11:1145 (2020)

PubMed id: 32123179

Nanobody-enabled monitoring of kappa opioid receptor states.

T.Che, J.English, B.E.Krumm, K.Kim, E.Pardon, R.H.J.Olsen, S.Wang, S.Zhang, J.F.Diberto, N.Sciaky, F.I.Carroll, J.Steyaert, D.Wacker, B.L.Roth.

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.