Single-molecule analysis of ligand efficacy in β2AR-G-protein activation
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Single-molecule analysis of ligand efficacy in β2AR-G-protein activation. / Gregorio, G. Glenn; Masureel, Matthieu; Hilger, Daniel; Terry, Daniel S.; Juette, Manuel; Zhao, Hong; Zhou, Zhou; Perez-Aguilar, Jose Manuel; Hauge, Maria; Mathiasen, Signe; Javitch, Jonathan A.; Weinstein, Harel; Kobilka, Brian K.; Blanchard, Scott C.
In: Nature, Vol. 547, No. 7661, 2017, p. 68–73.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Single-molecule analysis of ligand efficacy in β2AR-G-protein activation
AU - Gregorio, G. Glenn
AU - Masureel, Matthieu
AU - Hilger, Daniel
AU - Terry, Daniel S.
AU - Juette, Manuel
AU - Zhao, Hong
AU - Zhou, Zhou
AU - Perez-Aguilar, Jose Manuel
AU - Hauge, Maria
AU - Mathiasen, Signe
AU - Javitch, Jonathan A.
AU - Weinstein, Harel
AU - Kobilka, Brian K.
AU - Blanchard, Scott C.
PY - 2017
Y1 - 2017
N2 - G-protein-coupled receptor (GPCR)-mediated signal transduction is central to human physiology and disease intervention, yet the molecular mechanisms responsible for ligand-dependent signalling responses remain poorly understood. In class A GPCRs, receptor activation and G-protein coupling entail outward movements of transmembrane helix 6 (TM6). Here, using single-molecule fluorescence resonance energy transfer imaging, we examine TM6 movements in the β2 adrenergic receptor (β2AR) upon exposure to orthosteric ligands with different efficacies, in the absence and presence of the Gs heterotrimer. We show that partial and full agonists differentially affect TM6 motions to regulate the rate at which GDP-bound β2AR–Gs complexes are formed and the efficiency of nucleotide exchange leading to Gs activation. These data also reveal transient nucleotide-bound β2AR–Gs species that are distinct from known structures, and provide single-molecule perspectives on the allosteric link between ligand- and nucleotide-binding pockets that shed new light on the G-protein activation mechanism.
AB - G-protein-coupled receptor (GPCR)-mediated signal transduction is central to human physiology and disease intervention, yet the molecular mechanisms responsible for ligand-dependent signalling responses remain poorly understood. In class A GPCRs, receptor activation and G-protein coupling entail outward movements of transmembrane helix 6 (TM6). Here, using single-molecule fluorescence resonance energy transfer imaging, we examine TM6 movements in the β2 adrenergic receptor (β2AR) upon exposure to orthosteric ligands with different efficacies, in the absence and presence of the Gs heterotrimer. We show that partial and full agonists differentially affect TM6 motions to regulate the rate at which GDP-bound β2AR–Gs complexes are formed and the efficiency of nucleotide exchange leading to Gs activation. These data also reveal transient nucleotide-bound β2AR–Gs species that are distinct from known structures, and provide single-molecule perspectives on the allosteric link between ligand- and nucleotide-binding pockets that shed new light on the G-protein activation mechanism.
U2 - 10.1038/nature22354
DO - 10.1038/nature22354
M3 - Journal article
C2 - 28607487
VL - 547
SP - 68
EP - 73
JO - Nature
JF - Nature
SN - 0028-0836
IS - 7661
ER -
ID: 182486194