GPCR Engineering Yields High-Resolution Structural Insights into β <sub>2</sub> -Adrenergic Receptor Function
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- Daniel M. Rosenbaum
- Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
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- Vadim Cherezov
- Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
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- Michael A. Hanson
- Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
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- Søren G. F. Rasmussen
- Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
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- Foon Sun Thian
- Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
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- Tong Sun Kobilka
- Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
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- Hee-Jung Choi
- Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
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- Xiao-Jie Yao
- Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
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- William I. Weis
- Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
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- Raymond C. Stevens
- Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
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- Brian K. Kobilka
- Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
説明
<jats:p> The β <jats:sub>2</jats:sub> -adrenergic receptor (β <jats:sub>2</jats:sub> AR) 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 β <jats:sub>2</jats:sub> AR and to facilitate its crystallization, we engineered a β <jats:sub>2</jats:sub> AR fusion protein in which T4 lysozyme (T4L) replaces most of the third intracellular loop of the GPCR (“β <jats:sub>2</jats:sub> AR-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 β <jats:sub>2</jats:sub> AR-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. </jats:p>
収録刊行物
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- Science
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Science 318 (5854), 1266-1273, 2007-11-23
American Association for the Advancement of Science (AAAS)
