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PDBsum entry 6vi4
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Membrane protein
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PDB id
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6vi4
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References listed in PDB file
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Key reference
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Title
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Nanobody-Enabled monitoring of kappa opioid receptor states.
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Authors
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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.
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Ref.
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Nat Commun, 2020,
11,
1145.
[DOI no: ]
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PubMed id
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Abstract
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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.
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