Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Crystal structure of the human β2 adrenergic G-protein-coupled receptor


Structural analysis of G-protein-coupled receptors (GPCRs) for hormones and neurotransmitters has been hindered by their low natural abundance, inherent structural flexibility, and instability in detergent solutions. Here we report a structure of the human β2 adrenoceptor (β2AR), which was crystallized in a lipid environment when bound to an inverse agonist and in complex with a Fab that binds to the third intracellular loop. Diffraction data were obtained by high-brilliance microcrystallography and the structure determined at 3.4 Å/3.7 Å resolution. The cytoplasmic ends of the β2AR transmembrane segments and the connecting loops are well resolved, whereas the extracellular regions of the β2AR are not seen. The β2AR structure differs from rhodopsin in having weaker interactions between the cytoplasmic ends of transmembrane (TM)3 and TM6, involving the conserved E/DRY sequences. These differences may be responsible for the relatively high basal activity and structural instability of the β2AR, and contribute to the challenges in obtaining diffraction-quality crystals of non-rhodopsin GPCRs.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: Schematic diagram of the β 2 AR.
Figure 2: Structure of the β 2 AR365–Fab5 complex.
Figure 3: Comparison of β 2 AR and rhodopsin structures.
Figure 4: Side-chain interactions between Leu 272 and residues in TM3, TM5 and intracellular loop 2.


  1. 1

    Okada, T. et al. X-Ray diffraction analysis of three-dimensional crystals of bovine rhodopsin obtained from mixed micelles. J. Struct. Biol. 130, 73–80 (2000)

    CAS  Article  Google Scholar 

  2. 2

    Palczewski, K. et al. Crystal structure of rhodopsin: A G protein-coupled receptor. Science 289, 739–745 (2000)

    ADS  CAS  Article  Google Scholar 

  3. 3

    Okada, T. et al. Functional role of internal water molecules in rhodopsin revealed by X-ray crystallography. Proc. Natl Acad. Sci. USA 99, 5982–5987 (2002)

    ADS  CAS  Article  Google Scholar 

  4. 4

    Teller, D. C., Okada, T., Behnke, C. A., Palczewski, K. & Stenkamp, R. E. Advances in determination of a high-resolution three-dimensional structure of rhodopsin, a model of G-protein-coupled receptors (GPCRs). Biochemistry 40, 7761–7772 (2001)

    CAS  Article  Google Scholar 

  5. 5

    Okada, T. et al. The retinal conformation and its environment in rhodopsin in light of a new 2.2 A crystal structure. J. Mol. Biol. 342, 571–583 (2004)

    CAS  Article  Google Scholar 

  6. 6

    Li, J., Edwards, P. C., Burghammer, M., Villa, C. & Schertler, G. F. Structure of bovine rhodopsin in a trigonal crystal form. J. Mol. Biol. 343, 1409–1438 (2004)

    CAS  Article  Google Scholar 

  7. 7

    Standfuss, J. et al. Crystal structure of a thermally stable rhodopsin mutant. J. Mol. Biol. 372, 1179–1188 (2007)

    CAS  Article  Google Scholar 

  8. 8

    Lefkowitz, R. J. The superfamily of heptahelical receptors. Nature Cell Biol. 2, E133–E136 (2000)

    CAS  Article  Google Scholar 

  9. 9

    Strader, C. D., Sigal, I. S. & Dixon, R. A. Structural basis of β-adrenergic receptor function. FASEB J. 3, 1825–1832 (1989)

    CAS  Article  Google Scholar 

  10. 10

    Wieland, K., Zuurmond, H. M., Krasel, C., Ijzerman, A. P. & Lohse, M. J. Involvement of Asn-293 in stereospecific agonist recognition and in activation of the β2-adrenergic receptor. Proc. Natl Acad. Sci. USA 93, 9276–9281 (1996)

    ADS  CAS  Article  Google Scholar 

  11. 11

    Liapakis, G. et al. The forgotten serine. A critical role for Ser-2035.42 in ligand binding to and activation of the β2-adrenergic receptor. J. Biol. Chem. 275, 37779–37788 (2000)

    CAS  Article  Google Scholar 

  12. 12

    Strader, C. D., Candelore, M. R., Hill, W. S., Sigal, I. S. & Dixon, R. A. F. Identification of two serine residues involved in agonist activation of the β adrenergic receptor. J. Biol. Chem. 264, 13572–13578 (1989)

    CAS  Google Scholar 

  13. 13

    Ghanouni, P., Steenhuis, J. J., Farrens, D. L. & Kobilka, B. K. Agonist-induced conformational changes in the G-protein-coupling domain of the β2 adrenergic receptor. Proc. Natl Acad. Sci. USA 98, 5997–6002 (2001)

    ADS  CAS  Article  Google Scholar 

  14. 14

    Swaminath, G. et al. Sequential binding of agonists to the β2 adrenoceptor: kinetic evidence for intermediate conformational states. J. Biol. Chem. 279, 686–691 (2004)

    CAS  Article  Google Scholar 

  15. 15

    Ghanouni, P. et al. Functionally different agonists induce distinct conformations in the G protein coupling domain of the β2 adrenergic receptor. J. Biol. Chem. 276, 24433–24436 (2001)

    CAS  Article  Google Scholar 

  16. 16

    Swaminath, G. et al. Probing the β2 adrenoceptor binding site with catechol reveals differences in binding and activation by agonists and partial agonists. J.Biol. Chem. 280, 22165–22171 (2005)

    CAS  Article  Google Scholar 

  17. 17

    Yao, X. et al. Coupling ligand structure to specific conformational switches in the β2-adrenoceptor. Nature Chem. Biol. 2, 417–422 (2006)

    CAS  Article  Google Scholar 

  18. 18

    Kobilka, B. K. & Deupi, X. Conformational complexity of G-protein-coupled receptors. Trends Pharmacol. Sci. 28, 397–406 (2007)

    CAS  Article  Google Scholar 

  19. 19

    Kobilka, B. K. Amino and carboxyl terminal modifications to facilitate the production and purification of a G protein-coupled receptor. Anal. Biochem. 231, 269–271 (1995)

    CAS  Article  Google Scholar 

  20. 20

    Caron, M. G., Srinivasan, Y., Pitha, J., Kociolek, K. & Lefkowitz, R. J. Affinity chromatography of the β-adrenergic receptor. J. Biol. Chem. 254, 2923–2927 (1979)

    CAS  PubMed  Google Scholar 

  21. 21

    Kenakin, T. Ligand-selective receptor conformations revisited: the promise and the problem. Trends Pharmacol. Sci. 24, 346–354 (2003)

    CAS  Article  Google Scholar 

  22. 22

    Kobilka, B. K. G protein coupled receptor structure and activation. Biochim. Biophys. Acta 1768, 794–807 (2006)

    Article  Google Scholar 

  23. 23

    Gether, U. et al. Structural instability of a constitutively active G protein-coupled receptor. Agonist-independent activation due to conformational flexibility. J. Biol. Chem. 272, 2587–2590 (1997)

    CAS  Article  Google Scholar 

  24. 24

    Samama, P., Bond, R. A., Rockman, H. A., Milano, C. A. & Lefkowitz, R. J. Ligand-induced overexpression of a constitutively active β2-adrenergic receptor: Pharmacological creation of a phenotype in transgenic mice. Proc. Natl Acad. Sci. USA 94, 137–141 (1997)

    ADS  CAS  Article  Google Scholar 

  25. 25

    Granier, S. et al. Structure and conformational changes in the C-terminal domain of the β2-adrenoceptor: insights from fluorescence resonance energy transfer studies. J. Biol. Chem. 282, 13895–13905 (2007)

    CAS  Article  Google Scholar 

  26. 26

    Gether, U. & Kobilka, B. K. G protein-coupled receptors. II. Mechanism of agonist activation. J. Biol. Chem. 273, 17979–17982 (1998)

    CAS  Article  Google Scholar 

  27. 27

    Reiter, E. & Lefkowitz, R. J. GRKs and β-arrestins: roles in receptor silencing, trafficking and signaling. Trends Endocrinol. Metab. 17, 159–165 (2006)

    CAS  Article  Google Scholar 

  28. 28

    Day, P. W. et al. A monoclonal antibody for G protein coupled receptor crystallography. Nature Methods doi: 10.1038/nmeth1112 (21 October2007)

  29. 29

    Faham, S. et al. Crystallization of bacteriorhodopsin from bicelle formulations at room temperature. Protein Sci. 14, 836–840 (2005)

    CAS  Article  Google Scholar 

  30. 30

    Riekel, C., Burghammer, M. & Schertler, G. Protein crystallography microdiffraction. Curr. Opin. Struct. Biol. 15, 556–562 (2005)

    CAS  Article  Google Scholar 

  31. 31

    Faham, S. & Bowie, J. U. Bicelle crystallization: a new method for crystallizing membrane proteins yields a monomeric bacteriorhodopsin structure. J. Mol. Biol. 316, 1–6 (2002)

    CAS  Article  Google Scholar 

  32. 32

    Bulenger, S., Marullo, S. & Bouvier, M. Emerging role of homo- and heterodimerization in G-protein-coupled receptor biosynthesis and maturation. Trends Pharmacol. Sci. 26, 131–137 (2005)

    CAS  Article  Google Scholar 

  33. 33

    Whorton, M. R. et al. A monomeric G protein-coupled receptor isolated in a high-density lipoprotein particle efficiently activates its G protein. Proc. Natl Acad. Sci. USA 104, 7682–7687 (2007)

    ADS  CAS  Article  Google Scholar 

  34. 34

    Suryanarayana, S., Daunt, D. A., Von Zastrow, M. & Kobilka, B. K. A point mutation in the seventh hydrophobic domain of the α2 adrenergic receptor increases its affinity for a family of β receptor antagonists. J. Biol. Chem. 266, 15488–15492 (1991)

    CAS  PubMed  Google Scholar 

  35. 35

    Chelikani, P., Hornak, V., Eilers, M., Reeves, P. J., Smith, S. O., RajBhandary, U. L. & Khorana, H. G. Role of group-conserved residues in the helical core of β2-adrenergic receptor. Proc. Natl Acad. Sci. USA 104, 7027–7032 (2007)

    ADS  CAS  Article  Google Scholar 

  36. 36

    Tota, M. R. & Strader, C. D. Characterization of the binding domain of the β-adrenergic receptor with the fluorescent antagonist carazolol. Evidence for a buried ligand binding site. J. Biol. Chem. 265, 16891–16897 (1990)

    CAS  PubMed  Google Scholar 

  37. 37

    Ballesteros, J. A. et al. Activation of the β2-adrenergic receptor involves disruption of an ionic lock between the cytoplasmic ends of transmembrane segments 3 and 6. J. Biol. Chem. 276, 29171–29177 (2001)

    CAS  Article  Google Scholar 

  38. 38

    Scheer, A. et al. Mutational analysis of the highly conserved arginine within the Glu/Asp-Arg-Tyr motif of the α1b-adrenergic receptor: effects on receptor isomerization and activation. Mol. Pharmacol. 57, 219–231 (2000)

    CAS  PubMed  Google Scholar 

  39. 39

    Farrens, D. L., Altenbach, C., Yang, K., Hubbell, W. L. & Khorana, H. G. Requirement of rigid-body motion of transmembrane helices for light activation of rhodopsin. Science 274, 768–770 (1996)

    ADS  CAS  Article  Google Scholar 

  40. 40

    Salom, D. et al. Crystal structure of a photoactivated deprotonated intermediate of rhodopsin. Proc. Natl Acad. Sci. USA 103, 16123–16128 (2006)

    ADS  CAS  Article  Google Scholar 

  41. 41

    Samama, P., Cotecchia, S., Costa, T. & Lefkowitz, R. J. A mutation-induced activated state of the β2-adrenergic receptor. Extending the ternary complex model. J. Biol. Chem. 268, 4625–4636 (1993)

    CAS  PubMed  Google Scholar 

  42. 42

    Collaborative Computational Project. N. The CCP4 suite: programs for protein crystallography. Acta Crystallogr. D 50, 760–763 (1994)

  43. 43

    Otwinowski, Z. & Minor, W. Processing of x-ray diffraction data collected in oscillation mode. Methods Enzymol. 276, 307–326 (1997)

    CAS  Article  Google Scholar 

  44. 44

    Harris, L. J., Larson, S. B., Hasel, K. W. & McPherson, A. Refined structure of an intact IgG2a monoclonal antibody. Biochemistry 36, 1581–1597 (1997)

    CAS  Article  Google Scholar 

  45. 45

    McCoy, A. J. Solving structures of protein complexes by molecular replacement with Phaser. Acta Crystallogr. D 63, 32–41 (2007)

    CAS  Article  Google Scholar 

  46. 46

    Brünger, A. T. et al. Crystallography and NMR System (CNS): A new software system for macromolecular structure determination. Acta Crystallogr. D 54, 905–921 (1998)

    Article  Google Scholar 

Download references


This study was supported by the Lundbeck Foundation (S.G.F.R.), a National Institutes of Health Ruth L. Kirchstein NRSA grant (D.M.R.), a National Institute of General Medical Sciences grant (W.I.W.), a National Institute of Neurological Disorders and Stroke grant, the Mather Charitable Foundation, and a generous gift from Lundbeck (to B.K.K). G.F.X.S. was financially supported by a Human Frontier Science Project (HFSP) programme grant, a European Commission FP6 specific targeted research project and an ESRF long-term proposal. We thank R. Mackinnon and J. Bowie for advice, R. Stevens for help with early screening efforts, and J. Smith for arranging access to GM/CA-CAT at the APS. Use of the APS is supported by the US Department of Energy. GM/CA-CAT is funded by the US National Institutes of Cancer and General Medical Sciences. We thank X. Deupi and S. Granier for help with data collection. We thank D. Flot for his support at the ID 23.2 microfocus beamline at the European Synchrotron Radiation Facility.

Author Contributions S.G.F.R. performed final stages of β2AR purification, purified Mab5 and prepared Fab5. D.M.R. generated recombinant β2AR used for crystallography. Crystal screening and optimization were performed by S.G.F.R. and D.M.R. H.J.C. assisted with data collection at the Advanced Photon Source, processed all diffraction data and solved the structure of the β2AR–Fab5 complex. F.S.T. expressed β2AR in insect cells and, together with T.S.K., performed the initial stage of β2AR purification. T.S.K. prepared antibody 5. W.I.W. supervised and assisted with data collection at the Advanced Photon Source, and with data processing and structure determination. G.F.X.S. introduced B.K.K. to microfocus diffraction technology and supervised data collection at the European Synchrotron Radiation Facility. P.C.E. and M.B. assisted with data collection at the European Synchrotron Radiation Facility. R.S. and R.F.F. assisted with data collection at the Advanced Photon Source. V.R.P.R. performed the functional characterization of carazolol. B.K.K .was responsible for the overall project management and strategy, and assisted with β2AR purification, crystal harvesting and synchrotron data collection. B.K.K., W.I.W. and G.F.X.S. prepared the manuscript. All authors discussed the results and commented on the manuscript.

Author information



Corresponding author

Correspondence to Brian K. Kobilka.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

The file contains Supplementary Methods, Supplementary Table S1, Supplementary Figures S1-S5 and Legends. Supplementary Table S1 includes X-ray data collection and refinement statistics. Supplementary Figures S1-S3 and S5 show electron density maps for different regions of the crystal structure. Supplementary Figure S4 provides GTPγS binding data demonstrating that carazolol is a partial inverse agonist. This file was modified on 4 November 2007 to correct typographical errors. (PDF 10185 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Rasmussen, S., Choi, HJ., Rosenbaum, D. et al. Crystal structure of the human β2 adrenergic G-protein-coupled receptor. Nature 450, 383–387 (2007).

Download citation

Further reading


By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing