Structural flexibility of the G alpha s alpha-helical domain in the beta2-adrenoceptor Gs complex
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Structural flexibility of the G alpha s alpha-helical domain in the beta2-adrenoceptor Gs complex. / Westfield, Gerwin H; Rasmussen, Søren Gøgsig Faarup; Su, Min; Dutta, Somnath; DeVree, Brian T; Chung, Ka Young; Calinski, Diane; Velez-Ruiz, Gisselle; Oleskie, Austin N; Pardon, Els; Chae, Pil Seok; Liu, Tong; Li, Sheng; Woods, Virgil L; Steyaert, Jan; Kobilka, Brian K; Sunahara, Roger K; Skiniotis, Georgios.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 108, No. 38, 20.09.2011, p. 16086-91.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Structural flexibility of the G alpha s alpha-helical domain in the beta2-adrenoceptor Gs complex
AU - Westfield, Gerwin H
AU - Rasmussen, Søren Gøgsig Faarup
AU - Su, Min
AU - Dutta, Somnath
AU - DeVree, Brian T
AU - Chung, Ka Young
AU - Calinski, Diane
AU - Velez-Ruiz, Gisselle
AU - Oleskie, Austin N
AU - Pardon, Els
AU - Chae, Pil Seok
AU - Liu, Tong
AU - Li, Sheng
AU - Woods, Virgil L
AU - Steyaert, Jan
AU - Kobilka, Brian K
AU - Sunahara, Roger K
AU - Skiniotis, Georgios
PY - 2011/9/20
Y1 - 2011/9/20
N2 - The active-state complex between an agonist-bound receptor and a guanine nucleotide-free G protein represents the fundamental signaling assembly for the majority of hormone and neurotransmitter signaling. We applied single-particle electron microscopy (EM) analysis to examine the architecture of agonist-occupied β(2)-adrenoceptor (β(2)AR) in complex with the heterotrimeric G protein Gs (Gαsβγ). EM 2D averages and 3D reconstructions of the detergent-solubilized complex reveal an overall architecture that is in very good agreement with the crystal structure of the active-state ternary complex. Strikingly however, the α-helical domain of Gαs appears highly flexible in the absence of nucleotide. In contrast, the presence of the pyrophosphate mimic foscarnet (phosphonoformate), and also the presence of GDP, favor the stabilization of the α-helical domain on the Ras-like domain of Gαs. Molecular modeling of the α-helical domain in the 3D EM maps suggests that in its stabilized form it assumes a conformation reminiscent to the one observed in the crystal structure of Gαs-GTPγS. These data argue that the α-helical domain undergoes a nucleotide-dependent transition from a flexible to a conformationally stabilized state.
AB - The active-state complex between an agonist-bound receptor and a guanine nucleotide-free G protein represents the fundamental signaling assembly for the majority of hormone and neurotransmitter signaling. We applied single-particle electron microscopy (EM) analysis to examine the architecture of agonist-occupied β(2)-adrenoceptor (β(2)AR) in complex with the heterotrimeric G protein Gs (Gαsβγ). EM 2D averages and 3D reconstructions of the detergent-solubilized complex reveal an overall architecture that is in very good agreement with the crystal structure of the active-state ternary complex. Strikingly however, the α-helical domain of Gαs appears highly flexible in the absence of nucleotide. In contrast, the presence of the pyrophosphate mimic foscarnet (phosphonoformate), and also the presence of GDP, favor the stabilization of the α-helical domain on the Ras-like domain of Gαs. Molecular modeling of the α-helical domain in the 3D EM maps suggests that in its stabilized form it assumes a conformation reminiscent to the one observed in the crystal structure of Gαs-GTPγS. These data argue that the α-helical domain undergoes a nucleotide-dependent transition from a flexible to a conformationally stabilized state.
KW - Animals
KW - Crystallization
KW - Crystallography, X-Ray
KW - GTP-Binding Protein alpha Subunits, Gs
KW - Guanosine 5'-O-(3-Thiotriphosphate)
KW - Guanosine Diphosphate
KW - Guanosine Triphosphate
KW - Microscopy, Electron
KW - Models, Molecular
KW - Protein Binding
KW - Protein Conformation
KW - Protein Structure, Secondary
KW - Protein Structure, Tertiary
KW - Receptors, Adrenergic, beta-2
U2 - 10.1073/pnas.1113645108
DO - 10.1073/pnas.1113645108
M3 - Journal article
C2 - 21914848
VL - 108
SP - 16086
EP - 16091
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 38
ER -
ID: 120588171