Nanobody-enabled Monitoring of Kappa Opioid Receptor States

Overview

Journal Nat Commun

Specialty Biology

Date 2020 Mar 4

PMID 32123179

Citations 62

Authors

Tao Che

Justin English

Brian E Krumm

Kuglae Kim

Els Pardon

Reid H J Olsen

Sheng Wang

Shicheng Zhang

Jeffrey F DiBerto

Noah Sciaky

F Ivy Carroll

Jan Steyaert

Daniel Wacker

Bryan L Roth

Affiliations

Soon will be listed here.

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.

Citing Articles

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