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Originally published in Science Express on 25 October 2007
Science 23 November 2007:
Vol. 318. no. 5854, pp. 1266 - 1273
DOI: 10.1126/science.1150609
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Research Articles

GPCR Engineering Yields High-Resolution Structural Insights into β2-Adrenergic Receptor Function

Daniel M. Rosenbaum,1* Vadim Cherezov,2* Michael A. Hanson,2 Søren G. F. Rasmussen,1 Foon Sun Thian,1 Tong Sun Kobilka,1 Hee-Jung Choi,1,3 Xiao-Jie Yao,1 William I. Weis,1,3 Raymond C. Stevens,2{dagger} Brian K. Kobilka1{dagger}

The β2-adrenergic receptor (β2AR) 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 β2AR and to facilitate its crystallization, we engineered a β2AR fusion protein in which T4 lysozyme (T4L) replaces most of the third intracellular loop of the GPCR ("β2AR-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 β2AR-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.

1 Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
2 Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
3 Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.

* These authors contributed equally to this work.

{dagger} To whom correspondence should be addressed. E-mail: stevens{at}scripps.edu (R.C.S.); kobilka{at}stanford.edu (B.K.K.)

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