Structure of a nanobody-stabilized active state of the β(2) adrenoceptor

Søren Gøgsig Faarup Rasmussen; Hee-Jung Choi; Juan Jose Fung; Els Pardon; Paola Casarosa; Pil Seok Chae; Brian T Devree; Daniel M Rosenbaum; Foon Sun Thian; Tong Sun Kobilka; Andreas Schnapp; Ingo Konetzki; Roger K Sunahara; Samuel H Gellman; Alexander Pautsch; Jan Steyaert; William I Weis; Brian K Kobilka

doi:10.1038/nature09648

Structure of a nanobody-stabilized active state of the β(2) adrenoceptor

Søren Gøgsig Faarup Rasmussen, Hee-Jung Choi, Juan Jose Fung, Els Pardon, Paola Casarosa, Pil Seok Chae, Brian T Devree, Daniel M Rosenbaum, Foon Sun Thian, Tong Sun Kobilka, Andreas Schnapp, Ingo Konetzki, Roger K Sunahara, Samuel H Gellman, Alexander Pautsch, Jan Steyaert, William I Weis, Brian K Kobilka

Neuropharm and Genetics

1173 Citations (Scopus)

Abstract

G protein coupled receptors (GPCRs) exhibit a spectrum of functional behaviours in response to natural and synthetic ligands. Recent crystal structures provide insights into inactive states of several GPCRs. Efforts to obtain an agonist-bound active-state GPCR structure have proven difficult due to the inherent instability of this state in the absence of a G protein. We generated a camelid antibody fragment (nanobody) to the human β(2) adrenergic receptor (β(2)AR) that exhibits G protein-like behaviour, and obtained an agonist-bound, active-state crystal structure of the receptor-nanobody complex. Comparison with the inactive β(2)AR structure reveals subtle changes in the binding pocket; however, these small changes are associated with an 11 Å outward movement of the cytoplasmic end of transmembrane segment 6, and rearrangements of transmembrane segments 5 and 7 that are remarkably similar to those observed in opsin, an active form of rhodopsin. This structure provides insights into the process of agonist binding and activation.

Original language	English
Journal	Nature
Volume	469
Issue number	7329
Pages (from-to)	175-80
Number of pages	6
ISSN	0028-0836
DOIs	https://doi.org/10.1038/nature09648
Publication status	Published - 13 Jan 2011

Keywords

Adrenergic beta-2 Receptor Agonists
Animals
Binding Sites
Camelids, New World
Crystallography, X-Ray
Drug Inverse Agonism
Humans
Immunoglobulin Fragments
Ligands
Models, Molecular
Movement
Nanostructures
Opsins
Propanolamines
Protein Conformation
Protein Stability
Receptors, Adrenergic, beta-2
Viral Proteins

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Rasmussen, S. G. F., Choi, H-J., Fung, J. J., Pardon, E., Casarosa, P., Chae, P. S., Devree, B. T., Rosenbaum, D. M., Thian, F. S., Kobilka, T. S., Schnapp, A., Konetzki, I., Sunahara, R. K., Gellman, S. H., Pautsch, A., Steyaert, J., Weis, W. I., & Kobilka, B. K. (2011). Structure of a nanobody-stabilized active state of the β(2) adrenoceptor. Nature, 469(7329), 175-80. https://doi.org/10.1038/nature09648

Rasmussen, SGF, Choi, H-J, Fung, JJ, Pardon, E, Casarosa, P, Chae, PS, Devree, BT, Rosenbaum, DM, Thian, FS, Kobilka, TS, Schnapp, A, Konetzki, I, Sunahara, RK, Gellman, SH, Pautsch, A, Steyaert, J, Weis, WI & Kobilka, BK 2011, 'Structure of a nanobody-stabilized active state of the β(2) adrenoceptor', Nature, vol. 469, no. 7329, pp. 175-80. https://doi.org/10.1038/nature09648

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abstract = "G protein coupled receptors (GPCRs) exhibit a spectrum of functional behaviours in response to natural and synthetic ligands. Recent crystal structures provide insights into inactive states of several GPCRs. Efforts to obtain an agonist-bound active-state GPCR structure have proven difficult due to the inherent instability of this state in the absence of a G protein. We generated a camelid antibody fragment (nanobody) to the human β(2) adrenergic receptor (β(2)AR) that exhibits G protein-like behaviour, and obtained an agonist-bound, active-state crystal structure of the receptor-nanobody complex. Comparison with the inactive β(2)AR structure reveals subtle changes in the binding pocket; however, these small changes are associated with an 11 {\AA} outward movement of the cytoplasmic end of transmembrane segment 6, and rearrangements of transmembrane segments 5 and 7 that are remarkably similar to those observed in opsin, an active form of rhodopsin. This structure provides insights into the process of agonist binding and activation.",

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AU - Rasmussen, Søren Gøgsig Faarup

AU - Choi, Hee-Jung

AU - Fung, Juan Jose

AU - Pardon, Els

AU - Casarosa, Paola

AU - Chae, Pil Seok

AU - Devree, Brian T

AU - Rosenbaum, Daniel M

AU - Thian, Foon Sun

AU - Kobilka, Tong Sun

AU - Schnapp, Andreas

AU - Konetzki, Ingo

AU - Sunahara, Roger K

AU - Gellman, Samuel H

AU - Pautsch, Alexander

AU - Steyaert, Jan

AU - Weis, William I

AU - Kobilka, Brian K

PY - 2011/1/13

Y1 - 2011/1/13

N2 - G protein coupled receptors (GPCRs) exhibit a spectrum of functional behaviours in response to natural and synthetic ligands. Recent crystal structures provide insights into inactive states of several GPCRs. Efforts to obtain an agonist-bound active-state GPCR structure have proven difficult due to the inherent instability of this state in the absence of a G protein. We generated a camelid antibody fragment (nanobody) to the human β(2) adrenergic receptor (β(2)AR) that exhibits G protein-like behaviour, and obtained an agonist-bound, active-state crystal structure of the receptor-nanobody complex. Comparison with the inactive β(2)AR structure reveals subtle changes in the binding pocket; however, these small changes are associated with an 11 Å outward movement of the cytoplasmic end of transmembrane segment 6, and rearrangements of transmembrane segments 5 and 7 that are remarkably similar to those observed in opsin, an active form of rhodopsin. This structure provides insights into the process of agonist binding and activation.

AB - G protein coupled receptors (GPCRs) exhibit a spectrum of functional behaviours in response to natural and synthetic ligands. Recent crystal structures provide insights into inactive states of several GPCRs. Efforts to obtain an agonist-bound active-state GPCR structure have proven difficult due to the inherent instability of this state in the absence of a G protein. We generated a camelid antibody fragment (nanobody) to the human β(2) adrenergic receptor (β(2)AR) that exhibits G protein-like behaviour, and obtained an agonist-bound, active-state crystal structure of the receptor-nanobody complex. Comparison with the inactive β(2)AR structure reveals subtle changes in the binding pocket; however, these small changes are associated with an 11 Å outward movement of the cytoplasmic end of transmembrane segment 6, and rearrangements of transmembrane segments 5 and 7 that are remarkably similar to those observed in opsin, an active form of rhodopsin. This structure provides insights into the process of agonist binding and activation.

KW - Adrenergic beta-2 Receptor Agonists

KW - Animals

KW - Binding Sites

KW - Camelids, New World

KW - Crystallography, X-Ray

KW - Drug Inverse Agonism

KW - Humans

KW - Immunoglobulin Fragments

KW - Ligands

KW - Models, Molecular

KW - Movement

KW - Nanostructures

KW - Opsins

KW - Propanolamines

KW - Protein Conformation

KW - Protein Stability

KW - Receptors, Adrenergic, beta-2

KW - Viral Proteins

U2 - 10.1038/nature09648

DO - 10.1038/nature09648

M3 - Journal article

C2 - 21228869

SN - 0028-0836

VL - 469

SP - 175

EP - 180

JO - Nature

JF - Nature

IS - 7329

ER -

Structure of a nanobody-stabilized active state of the β(2) adrenoceptor

Abstract

Keywords

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