GPCR engineering yields high-resolution structural insights into beta2-adrenergic receptor function

TY - JOUR

T1 - GPCR engineering yields high-resolution structural insights into beta2-adrenergic receptor function

AU - Rosenbaum, Daniel M

AU - Cherezov, Vadim

AU - Hanson, Michael A

AU - Rasmussen, Søren Gøgsig Faarup

AU - Thian, Foon Sun

AU - Kobilka, Tong Sun

AU - Choi, Hee-Jung

AU - Yao, Xiao-Jie

AU - Weis, William I

AU - Stevens, Raymond C

AU - Kobilka, Brian K

PY - 2007/11/23

Y1 - 2007/11/23

N2 - The beta2-adrenergic receptor (beta2AR) is a well-studied prototype for heterotrimeric guanine nucleotide-binding protein (G protein)-coupled receptors (GPCRs) that respond to diffusible hormones and neurotransmitters. To overcome the structural flexibility of the beta2AR and to facilitate its crystallization, we engineered a beta2AR fusion protein in which T4 lysozyme (T4L) replaces most of the third intracellular loop of the GPCR ("beta2AR-T4L") and showed that this protein retains near-native pharmacologic properties. Analysis of adrenergic receptor ligand-binding mutants within the context of the reported high-resolution structure of beta2AR-T4L provides insights into inverse-agonist binding and the structural changes required to accommodate catecholamine agonists. Amino acids known to regulate receptor function are linked through packing interactions and a network of hydrogen bonds, suggesting a conformational pathway from the ligand-binding pocket to regions that interact with G proteins.

AB - The beta2-adrenergic receptor (beta2AR) is a well-studied prototype for heterotrimeric guanine nucleotide-binding protein (G protein)-coupled receptors (GPCRs) that respond to diffusible hormones and neurotransmitters. To overcome the structural flexibility of the beta2AR and to facilitate its crystallization, we engineered a beta2AR fusion protein in which T4 lysozyme (T4L) replaces most of the third intracellular loop of the GPCR ("beta2AR-T4L") and showed that this protein retains near-native pharmacologic properties. Analysis of adrenergic receptor ligand-binding mutants within the context of the reported high-resolution structure of beta2AR-T4L provides insights into inverse-agonist binding and the structural changes required to accommodate catecholamine agonists. Amino acids known to regulate receptor function are linked through packing interactions and a network of hydrogen bonds, suggesting a conformational pathway from the ligand-binding pocket to regions that interact with G proteins.

KW - Adrenergic beta-Agonists

KW - Adrenergic beta-Antagonists

KW - Amino Acid Sequence

KW - Bacteriophage T4

KW - Binding Sites

KW - Cell Line

KW - Cell Membrane

KW - Crystallization

KW - Crystallography, X-Ray

KW - Drug Inverse Agonism

KW - Humans

KW - Immunoglobulin Fab Fragments

KW - Ligands

KW - Models, Molecular

KW - Molecular Sequence Data

KW - Muramidase

KW - Propanolamines

KW - Protein Conformation

KW - Protein Structure, Secondary

KW - Protein Structure, Tertiary

KW - Receptors, Adrenergic, beta-2

KW - Recombinant Fusion Proteins

U2 - 10.1126/science.1150609

DO - 10.1126/science.1150609

M3 - Journal article

C2 - 17962519

SN - 0036-8075

VL - 318

SP - 1266

EP - 1273

JO - Science (New York, N.Y.)

JF - Science (New York, N.Y.)

IS - 5854

ER -