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 Citationer (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.

Originalsprog	Engelsk
Tidsskrift	Nature
Vol/bind	469
Udgave nummer	7329
Sider (fra-til)	175-80
Antal sider	6
ISSN	0028-0836
DOI	https://doi.org/10.1038/nature09648
Status	Udgivet - 13 jan. 2011

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10.1038/nature09648

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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, bind 469, nr. 7329, s. 175-80. https://doi.org/10.1038/nature09648

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title = "Structure of a nanobody-stabilized active state of the β(2) adrenoceptor",

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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author = "Rasmussen, {S{\o}ren G{\o}gsig Faarup} and Hee-Jung Choi and Fung, {Juan Jose} and Els Pardon and Paola Casarosa and Chae, {Pil Seok} and Devree, {Brian T} and Rosenbaum, {Daniel M} and Thian, {Foon Sun} and Kobilka, {Tong Sun} and Andreas Schnapp and Ingo Konetzki and Sunahara, {Roger K} and Gellman, {Samuel H} and Alexander Pautsch and Jan Steyaert and Weis, {William I} and Kobilka, {Brian K}",

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T1 - Structure of a nanobody-stabilized active state of the β(2) adrenoceptor

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

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