Conformational changes in the G protein Gs induced by the β2 adrenergic receptor (original) (raw)

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Acknowledgements

We thank J.J.G. Tesmer and G. Skiniotis for discussions. This work was supported by an American Lung Association senior research training fellowship (RT-166882-N, to K.Y.C.), the Lundbeck Foundation (a junior group leader fellowship, to S.G.F.R.), a National Institute of General Medical Sciences (NIGMS) molecular biophysics training grant (GM008270, to B.T.D), a National Institute of Neural Disorders and Stroke grant (NS28471, to B.K.K.), a NHLBI grant (HL071078, to B.K.K.), the Mather Charitable Foundation (B.K.K.), NIGMS grants GM083118 (to B.K.K. and R.K.S.) and GM068603 (to R.K.S.), a Michigan Diabetes Research and Training Center grant, the National Institute of Diabetes and Digestive and Kidney Diseases (P60DK-20572, to R.K.S.), the University of Michigan Biological Sciences Scholars Program (R.K.S.), and by the NIH (grants AI076961, AI081982, AI2008031, CA118595, GM20501, GM066170, GM093325 and RR029388 (to V.L.W.)).

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Author notes

  1. Pil Seok Chae
    Present address: Present address: Department of Bionano Engineering, Hanyang University, Ansan 426-791, Korea.,
  2. Ka Young Chung, Søren G. F. Rasmussen and Tong Liu: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, 94305, California, USA
    Ka Young Chung, Søren G. F. Rasmussen & Brian K. Kobilka
  2. Department of Neuroscience and Pharmacology, The Panum Institute, University of Copenhagen, 2200 Copenhagen N, Denmark,
    Søren G. F. Rasmussen
  3. Department of Medicine, Biomedical Sciences Graduate Program and UCSD DXMS Proteomics Resource, University of California San Diego, La Jolla, 92023, California, USA
    Tong Liu, Sheng Li & Virgil L. Woods Jr
  4. Department of Pharmacology, University of Michigan Medical School, Ann Arbor, 48109, Michigan, USA
    Brian T. DeVree, Diane Calinski & Roger K. Sunahara
  5. Department of Chemistry, University of Wisconsin, Madison, 53706, Wisconsin, USA
    Pil Seok Chae

Authors

  1. Ka Young Chung
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  2. Søren G. F. Rasmussen
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  3. Tong Liu
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  4. Sheng Li
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  5. Brian T. DeVree
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  6. Pil Seok Chae
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  7. Diane Calinski
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  8. Brian K. Kobilka
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  9. Virgil L. Woods Jr
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  10. Roger K. Sunahara
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Contributions

K.Y.C. prepared nucleotide-treated samples for DXMS, performed hydrogen–deuterium exchange of prepared samples, analysed mass spectrometry data, and performed cross-linking studies. S.G.F.R. performed the final stages of β2AR purification, assisted with β2AR and Gs protein virus production and expression in insect cell cultures, and worked out conditions to form, stabilize and purify the β2AR–Gs complex using agonist BI-167107 and detergent MNG-3. T.L. was responsible for the overall data analysis. S.L. ran prepared samples on the mass spectrometer. B.T.D. managed Gs heterotrimer subunit virus production and titration, expressed and purified Gs protein, and, with K.Y.C. and T.L., analysed mass spectrometry data. P.S.C. provided MNG-3 detergent for stabilization of the β2AR–Gs complex. D.C. assisted with Gs heterotrimer expression and purification. B.K.K. was responsible for overall project strategy and management, and assisted with manuscript preparation. V.L.W. oversaw design, execution and data analysis of DXMS experiments, and assisted with manuscript preparation. R.K.S. supervised Gs protein production, provided ideas and insights into Gs structure and function, and wrote the manuscript.

Corresponding authors

Correspondence toBrian K. Kobilka, Virgil L. Woods Jr or Roger K. Sunahara.

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The authors declare no competing financial interests.

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Chung, K., Rasmussen, S., Liu, T. et al. Conformational changes in the G protein Gs induced by the β2 adrenergic receptor.Nature 477, 611–615 (2011). https://doi.org/10.1038/nature10488

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