Allosteric nanobodies reveal the dynamic range and diverse mechanisms of G-protein-coupled receptor activation
Open accessAbstract
G-protein-coupled receptors (GPCRs) modulate many physiological processes by transducing a variety of extracellular cues into intracellular responses. Ligand binding to an extracellular orthosteric pocket propagates conformational change to the receptor cytosolic region to promote binding and activation of downstream signalling effectors such as G proteins and β-arrestins. It is well known that different agonists can share the same binding pocket but evoke unique receptor conformations leading to a wide range of downstream responses (‘efficacy’). Furthermore, increasing biophysical evidence, primarily using the β2-adrenergic receptor (β2AR) as a model system, supports the existence of multiple active and inactive conformational states. However, how agonists with varying efficacy modulate these receptor states to initiate cellular responses is not well understood. Here we report stabilization of two distinct β2AR conformations using single domain camelid antibodies (nanobodies)—a previously described positive allosteric nanobody (Nb80) and a newly identified negative allosteric nanobody (Nb60). We show that Nb60 stabilizes a previously unappreciated low-affinity receptor state which corresponds to one of two inactive receptor conformations as delineated by X-ray crystallography and NMR spectroscopy. We find that the agonist isoprenaline has a 15,000-fold higher affinity for β2AR in the presence of Nb80 compared to the affinity of isoprenaline for β2AR in the presence of Nb60, highlighting the full allosteric range of a GPCR. Assessing the binding of 17 ligands of varying efficacy to the β2AR in the absence and presence of Nb60 or Nb80 reveals large ligand-specific effects that can only be explained using an allosteric model which assumes equilibrium amongst at least three receptor states. Agonists generally exert efficacy by stabilizing the active Nb80-stabilized receptor state (R80). In contrast, for a number of partial agonists, both stabilization of R80 and destabilization of the inactive, Nb60-bound state (R60) contribute to their ability to modulate receptor activation. These data demonstrate that ligands can initiate a wide range of cellular responses by differentially stabilizing multiple receptor states.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
The absence of the L chain means that nanobody CDRs are distinct from those of antibodies in both sequence and structure, and as a result nanobodies are able to bind antibody-inaccessible epitopes in enzyme active sites, viral capsids and G protein coupled receptors.
“The absence of the L chain means that the nanobody’s CDRs are distinct from those of antibodies in both sequence and structure, and as a result nanobodies are able to bind antibody-inaccessible epitopes in enzyme active sites, viral capsids and G protein coupled receptors [233, 234].”
From Computational approaches to therapeutic antibody design: established methods and emerging trends (2020) · cites this paper as [233]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
[20] Processing of X-ray diffraction data collected in oscillation mode
Otwinowski, Z., & Minor, W. (1997) · Methods in Enzymology · doi:10.1016/s0076-6879(97)76066-x · Cited by 33,475 · see citing works
<i>Coot</i> : model-building tools for molecular graphics
Emsley, P., & Cowtan, K. (2004) · Acta Crystallographica Section D Biological Crystallography · doi:10.1107/s0907444904019158 · Cited by 30,571 · see citing works
Colowick, S. P., Joshi, M. D., Jagannathan, & KAPLAN, N. O. (1966) · Cited by 26,137 · see citing works
<i>PHENIX</i> : a comprehensive Python-based system for macromolecular structure solution
Adams, P. D., Afonine, P. V., Bunkóczi, G., Chen, V. B., Davis, I. W., Echols, N., Headd, J. J., Hung, L.-W., Kapral, G. J., Grosse-Kunstleve, R. W., et al. (2010) · Acta Crystallographica Section D Biological Crystallography · doi:10.1107/s0907444909052925 · Cited by 23,716 · see citing works
<i>Phaser</i> crystallographic software
McCoy, A. J., Grosse-Kunstleve, R. W., Adams, P. D., Winn, M. D., Storoni, L. C., & Read, R. J. (2007) · Journal of Applied Crystallography · doi:10.1107/s0021889807021206 · Cited by 20,612 · see citing works
<i>MolProbity</i> : all-atom structure validation for macromolecular crystallography
Chen, V. B., Arendall, W. B., Headd, J. J., Keedy, D. A., Immormino, R. M., Kapral, G. J., Murray, L. W., Richardson, J. S., & Richardson, D. C. (2010) · Acta Crystallographica Section D Biological Crystallography · doi:10.1107/s0907444909042073 · Cited by 14,115 · see citing works
Modified Spin-Echo Method for Measuring Nuclear Relaxation Times
Meiboom, S., & Gill, D. (1958) · Review of Scientific Instruments · doi:10.1063/1.1716296 · Cited by 6,001 · see citing works
Crystal structure of the β2 adrenergic receptor–Gs protein complex
Rasmussen, S. G. F., DeVree, B. T., Zou, Y., Kruse, A. C., Chung, K. Y., Kobilka, T. S., Thian, F. S., Chae, P. S., Pardon, E., Calinski, D., et al. (2011) · Nature · doi:10.1038/nature10361 · Cited by 3,009 · see citing works
Self-citationBallesteros, J. A., & Weinstein, H. (1995) · Methods in neurosciences · doi:10.1016/s1043-9471(05)80049-7 · Cited by 3,001 · see citing works
Structure of a nanobody-stabilized active state of the β2 adrenoceptor
Rasmussen, S. G. F., Choi, H., Fung, J. J., Pardon, E., Casarosa, P., Chae, P. S., DeVree, B. T., Rosenbaum, D. M., Thian, F. S., Kobilka, T. S., et al. (2011) · Nature · doi:10.1038/nature09648 · Cited by 1,663 · see citing works
Self-citationLéan, A. D., Stadel, J. M., & Lefkowitz, R. J. (1980) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(20)79672-9 · Cited by 1,619 · see citing works
Self-citationSamama, P., Cotecchia, S., Costa, T., & Lefkowitz, R. J. (1993) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)53442-6 · Cited by 1,256 · see citing works
Self-citationColquhoun, D. M. (1998) · British Journal of Pharmacology · doi:10.1038/sj.bjp.0702164 · Cited by 893 · see citing works
The Dynamic Process of β2-Adrenergic Receptor Activation
Nygaard, R., Zou, Y., Dror, R. O., Mildorf, T. J., Arlow, D. H., Manglik, A., Pan, A. C., Liu, C. W., Fung, J. J., Bokoch, M. P., et al. (2013) · Cell · doi:10.1016/j.cell.2013.01.008 · Cited by 797 · see citing works
Self-citationBiased Signaling Pathways in β 2 -Adrenergic Receptor Characterized by 19 F-NMR
Liu, J. J., Horst, R., Katritch, V., Stevens, R. C., & Kurt Wüthrich, A. (2012) · Science · doi:10.1126/science.1215802 · Cited by 697 · see citing works
Whorton, M. R., Bokoch, M. P., Rasmussen, S. G. F., Huang, B., Zare, R. N., Kobilka, B. K., & Sunahara, R. K. (2007) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0611448104 · Cited by 671 · see citing works
Self-citationStructural Insights into the Dynamic Process of β2-Adrenergic Receptor Signaling
Manglik, A., Kim, T. H., Masureel, M., Altenbach, C., Yang, Z., Hilger, D., Lerch, M. T., Kobilka, T. S., Thian, F. S., Hubbell, W. L., et al. (2015) · Cell · doi:10.1016/j.cell.2015.04.043 · Cited by 667 · see citing works
Self-citationBallesteros, J. A., Jensen, A. D., Liapakis, G., Rasmussen, S. G. F., Shi, L., Gether, U., & Javitch, J. A. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m103747200 · Cited by 619 · see citing works
Adrenaline-activated structure of β2-adrenoceptor stabilized by an engineered nanobody
Ring, A. M., Manglik, A., Kruse, A. C., Enos, M. D., Weis, W. I., García, K. C., & Kobilka, B. K. (2013) · Nature · doi:10.1038/nature12572 · Cited by 491 · see citing works
Self-citationQuantifying Ligand Bias at Seven-Transmembrane Receptors
Rajagopal, S., Ahn, S., Rominger, D. H., Gowen-MacDonald, W., Lam, C. M., DeWire, S. M., Violin, J. D., & Lefkowitz, R. J. (2011) · Molecular Pharmacology · doi:10.1124/mol.111.072801 · Cited by 391 · see citing works
Self-citationDror, R. O., Arlow, D. H., Borhani, D. W., Jensen, M. Ø., Piana, S., & Shaw, D. E. (2009) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0811065106 · Cited by 304 · see citing works
G-protein-coupled receptor inactivation by an allosteric inverse-agonist antibody
Hino, T., Arakawa, T., Iwanari, H., Yurugi-Kobayashi, T., Ikeda-Suno, C., Nakada-Nakura, Y., Kusano‐Arai, O., Weyand, S. N., Shimamura, T., Nomura, N., et al. (2012) · Nature · doi:10.1038/nature10750 · Cited by 297 · see citing works
Multiple ligand-specific conformations of the β2-adrenergic receptor
Kahsai, A. W., Xiao, K., Rajagopal, S., Ahn, S., Shukla, A. K., Sun, J., Oas, T. G., & Lefkowitz, R. J. (2011) · Nature Chemical Biology · doi:10.1038/nchembio.634 · Cited by 250 · see citing works
Self-citationKofuku, Y., Ueda, T., Okude, J., Shiraishi, Y., Kondo, K., Maeda, M., Tsujishita, H., & Shimada, I. (2012) · Nature Communications · doi:10.1038/ncomms2046 · Cited by 196 · see citing works
Moukhametzianov, R., Warne, T., Edwards, P. C., Serrano‐Vega, M. J., Leslie, A. G. W., Tate, C. G., & Schertler, G. F. X. (2011) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1100185108 · Cited by 176 · see citing works
Ehlert, F. J. (1985) · Molecular Pharmacology · doi:10.1016/s0026-895x(25)14182-5 · Cited by 168 · see citing works
Kobilka, B. K. (1995) · Analytical Biochemistry · doi:10.1006/abio.1995.1533 · Cited by 166 · see citing works
Self-citationWREGGETT, K. A., & Léan, A. D. (1984) · Molecular Pharmacology · doi:10.1016/s0026-895x(25)15078-5 · Cited by 151 · see citing works
The Dynamic Process of b 2 -Adrenergic Receptor Activation
Nygaard, R., Zou, Y., Dror, R. O., Mildorf, T. J., Arlow, D. H., Manglik, A., Pan, A. C., Liu, C. W., Bokoch, M. P., Thian, F. S., et al. (2013) · Cited by 146 · see citing works
Self-citationStaus, D. P., Wingler, L. M., Strachan, R. T., Rasmussen, S. G. F., Pardon, E., Ahn, S., Steyaert, J., Kobilka, B. K., & Lefkowitz, R. J. (2013) · Molecular Pharmacology · doi:10.1124/mol.113.089516 · Cited by 142 · see citing works
Self-citationA three-stage biophysical screening cascade for fragment-based drug discovery
Mashalidis, E. H., Sledz, P., Lang, S., & Abell, C. (2013) · Nature Protocols · doi:10.1038/nprot.2013.130 · Cited by 136 · see citing works
Strachan, R. T., Sun, J., Rominger, D. H., Violin, J. D., Ahn, S., Thomsen, A. R. B., Zhu, X., Kleist, A. B., Costa, T., & Lefkowitz, R. J. (2014) · Journal of Biological Chemistry · doi:10.1074/jbc.m114.548131 · Cited by 135 · see citing works
Self-citationThe quantitative analysis of drug–receptor interactions: a short history
Colquhoun, D. M. (2006) · Trends in Pharmacological Sciences · doi:10.1016/j.tips.2006.01.008 · Cited by 130 · see citing works
Tracing an allosteric pathway regulating the activity of the HslV protease
Shi, L., & Kay, L. E. (2014) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1318476111 · Cited by 82 · see citing works
Onaran, H. O., Rajagopal, S., & Costa, T. (2014) · Trends in Pharmacological Sciences · doi:10.1016/j.tips.2014.09.010 · Cited by 45 · see citing works
Self-citationAllosteric Coupling and Conformational Fluctuations in Proteins
Onaran, H. O., & Costa, T. (2009) · Current Protein and Peptide Science · doi:10.2174/138920309787847644 · Cited by 21 · see citing works
Self-citationVezzi, V., Onaran, H. O., Molinari, P., Guerrini, R., Balboni, G., Calò, G., & Costa, T. (2013) · Journal of Biological Chemistry · doi:10.1074/jbc.m113.474452 · Cited by 19 · see citing works
Self-citationComplex ligand-protein systems: a globally convergent iterative method for the n × m case
Pradines, J., Hasty, J., & Pakdaman, K. (2001) · Journal of Mathematical Biology · doi:10.1007/s002850100086 · Cited by 8 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
The Molecular Basis of G Protein–Coupled Receptor Activation
Weis, W. I., & Kobilka, B. K. (2018) · Annual Review of Biochemistry · doi:10.1146/annurev-biochem-060614-033910 · Cited by 1,227 · see citing works
Self-citationGPCR Dynamics: Structures in Motion
Latorraca, N. R., Venkatakrishnan, A., & Dror, R. O. (2016) · Chemical Reviews · doi:10.1021/acs.chemrev.6b00177 · Cited by 843 · see citing works
Biased signalling: from simple switches to allosteric microprocessors
Smith, J. S., Lefkowitz, R. J., & Rajagopal, S. (2018) · Nature Reviews Drug Discovery · doi:10.1038/nrd.2017.229 · Cited by 746 · see citing works
Self-citationCommon activation mechanism of class A GPCRs
Zhou, Q., Yang, D.-H., Wu, M., Guo, Y., Guo, W.-J., Zhong, L., Cai, X.-Q., Dai, A., Jang, W., Shakhnovich, E., et al. (2019) · eLife · doi:10.7554/elife.50279 · Cited by 491 · see citing works
How Ligands Illuminate GPCR Molecular Pharmacology
Wacker, D., Stevens, R. C., & Roth, B. L. (2017) · Cell · doi:10.1016/j.cell.2017.07.009 · Cited by 560 · see citing works
Yeast surface display platform for rapid discovery of conformationally selective nanobodies
McMahon, C., Baier, A. S., Pascolutti, R., Wegrecki, M., Zheng, S., Ong, J. X., Erlandson, S., Hilger, D., Rasmussen, S., Ring, A., et al. (2018) · Nature Structural & Molecular Biology · doi:10.1038/s41594-018-0028-6 · Cited by 493 · see citing works
Self-citationAn ultrapotent synthetic nanobody neutralizes SARS-CoV-2 by stabilizing inactive Spike
Schoof, M., Faust, B., Saunders, R. A., Sangwan, S., Rezelj, V. V., Hoppe, N., Boone, M., Billesbølle, C. B., Puchades, C., Azumaya, C. M., et al. (2020) · Science · doi:10.1126/science.abe3255 · Cited by 428 · see citing works
Self-citationMini G protein probes for active G protein–coupled receptors (GPCRs) in live cells
Wan, Q., Okashah, N., Inoue, A., Nehmé, R., Carpenter, B., Tate, C. G., & Lambert, N. A. (2018) · Journal of Biological Chemistry · doi:10.1074/jbc.ra118.001975 · Cited by 391 · see citing works
Structural insights into G-protein-coupled receptor allostery
Thal, D. M., Glukhova, A., Sexton, P. M., & Christopoulos, A. (2018) · Nature · doi:10.1038/s41586-018-0259-z · Cited by 320 · see citing works
Biased Receptor Signaling in Drug Discovery
Kenakin, T. (2019) · Pharmacological Reviews · doi:10.1124/pr.118.016790 · Cited by 302 · see citing works
Structure of CC chemokine receptor 2 with orthosteric and allosteric antagonists
Zheng, Y., Qin, L., Zacarı́as, N. V. O., de Vries, H., Han, G. W., Gustavsson, M. B., Dabros, M., Zhao, C., Cherney, R. J., Carter, P., et al. (2016) · Nature · doi:10.1038/nature20605 · Cited by 288 · see citing works
A Genetically Encoded Biosensor Reveals Location Bias of Opioid Drug Action
Stoeber, M., Jullié, D., Lobingier, B. T., Laeremans, T., Steyaert, J., Schiller, P. W., Manglik, A., & von Zastrow, M. (2018) · Neuron · doi:10.1016/j.neuron.2018.04.021 · Cited by 288 · see citing works
Self-citationSynthetic single domain antibodies for the conformational trapping of membrane proteins
Zimmermann, I., Egloff, P., Hutter, C. A., Arnold, F. M., Stohler, P., Bocquet, N., Hug, M. N., Huber, S., Siegrist, M., Hetemann, L., et al. (2018) · eLife · doi:10.7554/elife.34317 · Cited by 283 · see citing works
GPCR drug discovery: integrating solution NMR data with crystal and cryo-EM structures
Shimada, I., Ueda, T., Kofuku, Y., Eddy, M. T., & Wüthrich, K. (2018) · Nature Reviews Drug Discovery · doi:10.1038/nrd.2018.180 · Cited by 282 · see citing works
Comparative analysis of nanobody sequence and structure data
Mitchell, L. S., & Colwell, L. J. (2018) · Proteins Structure Function and Bioinformatics · doi:10.1002/prot.25497 · Cited by 233 · see citing works
Foster, D. J., & Conn, P. J. (2017) · Neuron · doi:10.1016/j.neuron.2017.03.016 · Cited by 227 · see citing works
Computational approaches to therapeutic antibody design: established methods and emerging trends
Norman, R. A., Ambrosetti, F., Bonvin, A. M. J. J., Colwell, L. J., Kelm, S., Kumar, S., & Krawczyk, K. (2020) · Briefings in Bioinformatics · doi:10.1093/bib/bbz095 · Cited by 219 · see citing works
“The absence of the L chain means that the nanobody’s CDRs are distinct from those of antibodies in both sequence and structure, and as a result nanobodies are able to bind antibody-inaccessible epitopes in enzyme active sites, viral capsids and G protein coupled receptors [233, 234].”
BackgroundConformational Basis of G Protein-Coupled Receptor Signaling Versatility
Wingler, L. M., & Lefkowitz, R. J. (2020) · Trends in Cell Biology · doi:10.1016/j.tcb.2020.06.002 · Cited by 222 · see citing works
Self-citationAllosteric Coupling of Drug Binding and Intracellular Signaling in the A2A Adenosine Receptor
Eddy, M. T., Lee, M.-Y., Gao, Z., White, K. L., Didenko, T., Horst, R., Audet, M., Stańczak, P., McClary, K. M., Han, G. W., et al. (2017) · Cell · doi:10.1016/j.cell.2017.12.004 · Cited by 212 · see citing works
Mechanism of intracellular allosteric β2AR antagonist revealed by X-ray crystal structure
Liu, X., Ahn, S., Kahsai, A. W., Meng, K.-C., Latorraca, N. R., Pani, B., Venkatakrishnan, A., Masoudi, A., Weis, W. I., Dror, R. O., et al. (2017) · Nature · doi:10.1038/nature23652 · Cited by 203 · see citing works
Self-citationBeghein, E., & Gettemans, J. (2017) · Frontiers in Immunology · doi:10.3389/fimmu.2017.00771 · Cited by 190 · see citing works
Structural determinants of 5-HT2B receptor activation and biased agonism
McCorvy, J. D., Wacker, D., Wang, S., Agegnehu, B., Liu, J., Lansu, K., Tribo, A. R., Olsen, R. H. J., Che, T., Jin, J. J., et al. (2018) · Nature Structural & Molecular Biology · doi:10.1038/s41594-018-0116-7 · Cited by 174 · see citing works
Allosteric “beta-blocker” isolated from a DNA-encoded small molecule library
Ahn, S., Kahsai, A. W., Pani, B., Wang, Q.-T., Zhao, S., Wall, A. L., Strachan, R. T., Staus, D. P., Wingler, L. M., Sun, L., et al. (2017) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1620645114 · Cited by 162 · see citing works
Self-citationHsp70 chaperone: a master player in protein homeostasis
Fernández-Fernández, M. R., & Valpuesta, J. (2018) · F1000Research · doi:10.12688/f1000research.15528.1 · Cited by 162 · see citing works
Ballante, F., Kooistra, A. J., Kampen, S., de Graaf, C., & Carlsson, J. (2021) · Pharmacological Reviews · doi:10.1124/pharmrev.120.000246 · Cited by 160 · see citing works
Lawson, A. D. G., MacCoss, M., & Heer, J. P. (2017) · Journal of Medicinal Chemistry · doi:10.1021/acs.jmedchem.7b01120 · Cited by 154 · see citing works
Delineating the conformational landscape of the adenosine A2A receptor during G protein coupling
Huang, S. K., Pandey, A. N., Tran, D. P., Villanueva, N., Kitao, A., Sunahara, R. K., Sljoka, A., & Prosser, R. S. (2021) · Cell · doi:10.1016/j.cell.2021.02.041 · Cited by 148 · see citing works
Self-citationRecombinant expression of nanobodies and nanobody-derived immunoreagents
de Marco, A. (2020) · Protein Expression and Purification · doi:10.1016/j.pep.2020.105645 · Cited by 137 · see citing works
Nanobody-enabled monitoring of kappa opioid receptor states
Che, T., English, J. G., Krumm, B. E., Kim, K., Pardon, E., Olsen, R. H. J., Wang, S., Zhang, S., DiBerto, J. F., Sciaky, N., et al. (2020) · Nature Communications · doi:10.1038/s41467-020-14889-7 · Cited by 134 · see citing works
Self-citationRapid generation of potent antibodies by autonomous hypermutation in yeast
Wellner, A., McMahon, C. T., Gilman, M. S. A., Clements, J. R., Clark, S. A., Nguyen, K. M., Ho, M. H., Hu, V. J., Shin, J.-E., Feldman, J. E., et al. (2021) · Nature Chemical Biology · doi:10.1038/s41589-021-00832-4 · Cited by 128 · see citing works
Self-citationMechanism of β 2 AR regulation by an intracellular positive allosteric modulator
Liu, X., Masoudi, A., Kahsai, A. W., Huang, L.-Y., Pani, B., Staus, D. P., Shim, P. J., Hirata, K., Simhal, R. K., Schwalb, A. M., et al. (2019) · Science · doi:10.1126/science.aaw8981 · Cited by 128 · see citing works
Self-citationThe Endocannabinoid System as a Target in Cancer Diseases: Are We There Yet?
Moreno, E., Cavic, M. R., Krivokuća, A., Casadó, V., & Canela, E. I. (2019) · Frontiers in Pharmacology · doi:10.3389/fphar.2019.00339 · Cited by 126 · see citing works
Structural Basis for G Protein–Coupled Receptor Signaling
Erlandson, S. C., McMahon, C. T., & Kruse, A. C. (2018) · Annual Review of Biophysics · doi:10.1146/annurev-biophys-070317-032931 · Cited by 126 · see citing works
Self-citationAn online resource for GPCR structure determination and analysis
Munk, C., Mutt, E., Ísberg, V., Nikolajsen, L. F., Bibbe, J. M., Flock, T., Hanson, M. A., Stevens, R. C., Deupí, X., & Gloriam, D. E. I. (2019) · Nature Methods · doi:10.1038/s41592-018-0302-x · Cited by 125 · see citing works
Haloperidol bound D2 dopamine receptor structure inspired the discovery of subtype selective ligands
Fan, L., Tan, L., Chen, Z., Qi, J., Nie, F., Luo, Z., Cheng, J., & Wang, S. (2020) · Nature Communications · doi:10.1038/s41467-020-14884-y · Cited by 120 · see citing works
Single-molecule analysis reveals agonist-specific dimer formation of µ-opioid receptors
Møller, J., Işbilir, A., Sungkaworn, T., Osberg, B., Karathanasis, C., Sunkara, V., Grushevskyi, E. O., Böck, A., Annibale, P., Heilemann, M., et al. (2020) · Nature Chemical Biology · doi:10.1038/s41589-020-0566-1 · Cited by 120 · see citing works
Molecular basis for high-affinity agonist binding in GPCRs
Warne, T., Edwards, P. C., Dore, A. S., Leslie, A. G. W., & Tate, C. G. (2019) · Science · doi:10.1126/science.aau5595 · Cited by 119 · see citing works
G Protein–Coupled Receptors in Asthma Therapy: Pharmacology and Drug Action
Wendell, S. G., Fan, H., & Zhang, C. (2019) · Pharmacological Reviews · doi:10.1124/pr.118.016899 · Cited by 117 · see citing works
Allosteric drugs and mutations: chances, challenges, and necessity
Guarnera, E., & Berezovsky, I. N. (2020) · Current Opinion in Structural Biology · doi:10.1016/j.sbi.2020.01.010 · Cited by 117 · see citing works
Lee, Y., Basith, S., & Choi, S. (2017) · Journal of Medicinal Chemistry · doi:10.1021/acs.jmedchem.6b01453 · Cited by 116 · see citing works
Formation of pH‐Resistant Monodispersed Polymer–Lipid Nanodiscs
Ravula, T., Hardin, N. Z., Ramadugu, S. K., Cox, S. J., & Ramamoorthy, A. (2017) · Angewandte Chemie International Edition · doi:10.1002/anie.201712017 · Cited by 116 · see citing works
The role of structural dynamics in GPCR‐mediated signaling
Hilger, D. (2021) · FEBS Journal · doi:10.1111/febs.15841 · Cited by 116 · see citing works
Structures of full-length glycoprotein hormone receptor signalling complexes
Duan, J., Xu, P., Cheng, X., Mao, C., Croll, T. I., He, X., Shi, J., Luan, X., Yin, W., You, E., et al. (2021) · Nature · doi:10.1038/s41586-021-03924-2 · Cited by 114 · see citing works
Li, Z., Zhang, W., Zhang, Q., Li, P., & Tang, X. (2023) · ACS Nano · doi:10.1021/acsnano.3c06930 · Cited by 110 · see citing works
Autoregulation of GPCR signalling through the third intracellular loop
Sadler, F. W., Ma, N., Ritt, M., Sharma, Y. K., Vaidehi, N., & Sivaramakrishnan, S. (2023) · Nature · doi:10.1038/s41586-023-05789-z · Cited by 106 · see citing works
Biased Agonism in Drug Discovery—Is It Too Soon to Choose a Path?
Michel, M. C., & Charlton, S. J. (2018) · Molecular Pharmacology · doi:10.1124/mol.117.110890 · Cited by 104 · see citing works
Gαi is required for carvedilol-induced β1 adrenergic receptor β-arrestin biased signaling
Wang, J., Hanada, K., Staus, D. P., Makara, M. A., Dahal, G. R., Chen, Q., Ahles, A., Engelhardt, S. H., & Rockman, H. A. (2017) · Nature Communications · doi:10.1038/s41467-017-01855-z · Cited by 105 · see citing works
Self-citationWhat Do Structures Tell Us About Chemokine Receptor Function and Antagonism?
Kufareva, I., Gustavsson, M. B., Zheng, Y., Stephens, B. S., & Handel, T. M. (2017) · Annual Review of Biophysics · doi:10.1146/annurev-biophys-051013-022942 · Cited by 101 · see citing works
VEGAS as a Platform for Facile Directed Evolution in Mammalian Cells
English, J. G., Olsen, R. H. J., Lansu, K., Patel, M., White, K., Cockrell, A. S., Singh, D., Strachan, R. T., Wacker, D., & Roth, B. L. (2019) · Cell · doi:10.1016/j.cell.2019.05.051 · Cited by 98 · see citing works
Self-citationExploring cellular biochemistry with nanobodies
Cheloha, R. W., Harmand, T. J., Wijne, C., Schwartz, T., & Ploegh, H. L. (2020) · Journal of Biological Chemistry · doi:10.1074/jbc.rev120.012960 · Cited by 92 · see citing works
Scholler, P., Névoltris, D., Bundel, D. D., Bossi, S., Moreno‐Delgado, D., Rovira, X., Møller, T. C., Moustaine, D. E., Mathieu, M., Blanc, É., et al. (2017) · Nature Communications · doi:10.1038/s41467-017-01489-1 · Cited by 94 · see citing works
Devost, D., Sleno, R., Pétrin, D., Zhang, A., Shinjo, Y., Okde, R., Aoki, J., Inoue, A., & Hébert, T. E. (2017) · Journal of Biological Chemistry · doi:10.1074/jbc.m116.763854 · Cited by 92 · see citing works
Nanobodies to study protein conformational states
Uchański, T., Pardon, E., & Steyaert, J. (2020) · Current Opinion in Structural Biology · doi:10.1016/j.sbi.2020.01.003 · Cited by 92 · see citing works
Self-citationStructural basis of species-selective antagonist binding to the succinate receptor
Haffke, M., Fehlmann, D., Rummel, G., Boivineau, J., Duckely, M., Gommermann, N., Cotesta, S., Sirockin, F., Freuler, F., Littlewood-Evans, A. J., et al. (2019) · Nature · doi:10.1038/s41586-019-1663-8 · Cited by 90 · see citing works
Nonantimicrobial Actions of Macrolides: Overview and Perspectives for Future Development
Kricker, J. A., Page, C. P., Gardarsson, F. R., Baldursson, Ó., Guðjónsson, Þ., & Parnham, M. J. (2021) · Pharmacological Reviews · doi:10.1124/pharmrev.121.000300 · Cited by 92 · see citing works
Defining a conformational ensemble that directs activation of PPARγ
Chrisman, I. M., Nemetchek, M. D., de Vera, I. M. S., Shang, J., Heidari, Z., Long, Y., Reyes-Caballero, H., Galindo‐Murillo, R., Cheatham, T. E., Blayo, A., et al. (2018) · Nature Communications · doi:10.1038/s41467-018-04176-x · Cited by 88 · see citing works
Ahn, S., Pani, B., Kahsai, A. W., Olsen, E. K., Husemoen, G., Vestergaard, M., Jin, L., Zhao, S., Wingler, L. M., Rambarat, P. K., et al. (2018) · Molecular Pharmacology · doi:10.1124/mol.118.111948 · Cited by 87 · see citing works
Self-citationLigand and G-protein selectivity in the κ-opioid receptor
Han, J., Zhang, J., Nazarova, A. L., Bernhard, S. M., Krumm, B. E., Zhao, L., Lam, J. H., Rangari, V. A., Majumdar, S., Nichols, D. E., et al. (2023) · Nature · doi:10.1038/s41586-023-06030-7 · Cited by 85 · see citing works
Energy Landscapes Reveal Agonist Control of G Protein-Coupled Receptor Activation via Microswitches
Fleetwood, O., Matricon, P. G., Carlsson, J., & Delemotte, L. (2020) · Biochemistry · doi:10.1021/acs.biochem.9b00842 · Cited by 84 · see citing works
G protein-coupled receptors: the evolution of structural insight
Gacasan, S. B., Baker, D. L., & Parrill, A. L. (2017) · AIMS Biophysics · doi:10.3934/biophy.2017.3.491 · Cited by 83 · see citing works
Shiraishi, Y., Natsume, M., Kofuku, Y., Imai, S., Nakata, K., Mizukoshi, T., Ueda, T., Iwaï, H., & Shimada, I. (2018) · Nature Communications · doi:10.1038/s41467-017-02632-8 · Cited by 79 · see citing works
Allosteric modulators targeting CNS muscarinic receptors
Böck, A., Schrage, R., & Mohr, K. (2017) · Neuropharmacology · doi:10.1016/j.neuropharm.2017.09.024 · Cited by 78 · see citing works
Allosteric coupling and biased agonism in G protein‐coupled receptors
Böck, A., & Bermúdez, M. (2021) · FEBS Journal · doi:10.1111/febs.15783 · Cited by 76 · see citing works
Allosteric modulation of G protein-coupled receptor signaling
Shen, S., Zhao, C. S., Wu, C., Sun, S., Li, Z., Yan, W. X., & Shao, Z. (2023) · Frontiers in Endocrinology · doi:10.3389/fendo.2023.1137604 · Cited by 75 · see citing works
Liu, X., Masoudi, A., Kahsai, A. W., Huang, L.-Y., Pani, B., Hirata, K., Ahn, S., Lefkowitz, R. J., & Kobilka, B. K. (2018) · Worldwide Protein Data Bank · doi:10.2210/pdb6n48/pdb · Cited by 75 · see citing works
Self-citationAminergic GPCR–Ligand Interactions: A Chemical and Structural Map of Receptor Mutation Data
Vass, M., Podlewska, S., de Esch, I. J. P., Bojarski, A. J., Leurs, R., Kooistra, A. J., & de Graaf, C. (2018) · Journal of Medicinal Chemistry · doi:10.1021/acs.jmedchem.8b00836 · Cited by 74 · see citing works
Molecular pharmacology of metabotropic receptors targeted by neuropsychiatric drugs
Roth, B. L. (2019) · Nature Structural & Molecular Biology · doi:10.1038/s41594-019-0252-8 · Cited by 68 · see citing works
New Insights into the Structure and Function of Class B1 GPCRs
Cary, B. P., Zhang, X., Cao, J., Johnson, R. M., Piper, S., Gerrard, E. J., Wootten, D. L., & Sexton, P. M. (2022) · Endocrine Reviews · doi:10.1210/endrev/bnac033 · Cited by 69 · see citing works
Recognition of methamphetamine and other amines by trace amine receptor TAAR1
Liu, H., Zheng, Y. Y., Wang, Y., Wang, Y., He, X., Xu, P., Huang, S., Yuan, Q., Zhang, X., Wang, L., et al. (2023) · Nature · doi:10.1038/s41586-023-06775-1 · Cited by 69 · see citing works
Human antibody-based chemically induced dimerizers for cell therapeutic applications
Hill, Z. B., Martinko, A. J., Nguyen, D., & Wells, J. A. (2017) · Nature Chemical Biology · doi:10.1038/nchembio.2529 · Cited by 67 · see citing works
PAM-Antagonists: A Better Way to Block Pathological Receptor Signaling?
Kenakin, T., & Strachan, R. T. (2018) · Trends in Pharmacological Sciences · doi:10.1016/j.tips.2018.05.001 · Cited by 67 · see citing works
Self-citationFSH for the Treatment of Male Infertility
Casarini, L., Crépieux, P., Reiter, É., Lazzaretti, C., Paradiso, E., Rochira, V., Brigante, G., Santi, D., & Simoni, M. (2020) · International Journal of Molecular Sciences · doi:10.3390/ijms21072270 · Cited by 70 · see citing works
Fleetwood, O., Carlsson, J., & Delemotte, L. (2021) · eLife · doi:10.7554/elife.60715 · Cited by 66 · see citing works
Grahl, A., Abiko, L. A., Isogai, S., Sharpe, T., & Grzesiek, S. (2020) · Nature Communications · doi:10.1038/s41467-020-15864-y · Cited by 64 · see citing works
Nanobodies: New avenues for imaging, stabilizing and modulating GPCRs
Groof, T. W. M. D., Bobkov, V., Heukers, R., & Smit, M. J. (2019) · Molecular and Cellular Endocrinology · doi:10.1016/j.mce.2019.01.021 · Cited by 62 · see citing works
Khamlichi, C. E., Reverchon, F., Hervouet, N., Blot, L., Reiter, É., & Morisset‐Lopez, S. (2019) · Molecules · doi:10.3390/molecules24030537 · Cited by 62 · see citing works
Structural basis for ion selectivity in TMEM175 K+ channels
Brunner, J. D., Jakob, R. P., Schulze, T., Neldner, Y., Moroni, A., Thiel, G., Maier, T., & Schenck, S. (2020) · eLife · doi:10.7554/elife.53683 · Cited by 60 · see citing works
Li, X., Anderson, M. D., Collin, D., Muegge, I., Wan, J., Brennan, D. L., Kugler, S. Z., Terenzio, D., Kennedy, C., Lin, S., et al. (2017) · Journal of Biological Chemistry · doi:10.1074/jbc.m117.789024 · Cited by 61 · see citing works
Nanobodies detecting and modulating GPCRs outside in and inside out
Heukers, R., Groof, T. W. M. D., & Smit, M. J. (2019) · Current Opinion in Cell Biology · doi:10.1016/j.ceb.2019.01.003 · Cited by 59 · see citing works
Enabling STD-NMR fragment screening using stabilized native GPCR: A case study of adenosine receptor
Igonet, S., Raingeval, C., Cecon, E., Pučić‐Baković, M., Lauc, G., Cala, O., Baranowski, M., Pérez, J., Jockers, R., Krimm, I., et al. (2018) · Scientific Reports · doi:10.1038/s41598-018-26113-0 · Cited by 58 · see citing works
Shpakov, A. O. (2023) · International Journal of Molecular Sciences · doi:10.3390/ijms24076187 · Cited by 56 · see citing works
Intracellular Receptor Modulation: Novel Approach to Target GPCRs
Zacarı́as, N. V. O., Lenselink, E. B., IJzerman, A. P., Handel, T. M., & Heitman, L. H. (2018) · Trends in Pharmacological Sciences · doi:10.1016/j.tips.2018.03.002 · Cited by 56 · see citing works
Miles, T. F., Spieß, K., Jude, K. M., Tsutsumi, N., Burg, J. S., Ingram, J. R., Waghray, D., Hjortø, G. M., Larsen, O., Ploegh, H. L., et al. (2018) · eLife · doi:10.7554/elife.35850 · Cited by 55 · see citing works
Duan, J., Xu, P., Zhang, H., Luan, X., Yang, J., He, X., Mao, C., Shen, D., Ji, Y., Cheng, X., et al. (2023) · Nature Communications · doi:10.1038/s41467-023-36170-3 · Cited by 54 · see citing works
Fluorescent probes for G-protein-coupled receptor drug discovery
Iliopoulos‐Tsoutsouvas, C., Kulkarni, R., Makriyannis, A., & Nikas, S. P. (2018) · Expert Opinion on Drug Discovery · doi:10.1080/17460441.2018.1518975 · Cited by 54 · see citing works
Stepwise activation of a class C GPCR begins with millisecond dimer rearrangement
Grushevskyi, E. O., Kukaj, T., Schmauder, R., Böck, A., Zabel, U., Schwabe, T., Benndorf, K., & Lohse, M. J. (2019) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1900261116 · Cited by 54 · see citing works
A Critical Analysis of Molecular Mechanisms Underlying Membrane Cholesterol Sensitivity of GPCRs
Jafurulla, M., Kumar, G. A., Rao, B. D., & Chattopadhyay, A. (2019) · Advances in experimental medicine and biology · doi:10.1007/978-3-030-04278-3_2 · Cited by 53 · see citing works
β 2 -Adrenergic Receptor Conformational Response to Fusion Protein in the Third Intracellular Loop
Eddy, M. T., Didenko, T., Stevens, R. C., & Kurt Wüthrich, A. (2016) · Structure · doi:10.1016/j.str.2016.09.015 · Cited by 52 · see citing works
How Carvedilol activates β2-adrenoceptors
Benkel, T., Zimmermann, M., Zeiner, J., Bravo, S., Merten, N., Lim, V., Matthees, E. S. F., Drube, J., Miess, E., Malan, D., et al. (2022) · Nature Communications · doi:10.1038/s41467-022-34765-w · Cited by 52 · see citing works
Measuring ligand efficacy at the mu-opioid receptor using a conformational biosensor
Livingston, K. E., Mahoney, J. P., Manglik, A., Sunahara, R. K., & Traynor, J. R. (2018) · eLife · doi:10.7554/elife.32499 · Cited by 51 · see citing works
Self-citationToward an understanding of the structural basis of allostery in muscarinic acetylcholine receptors
Burger, W. A. C., Sexton, P. M., Christopoulos, A., & Thal, D. M. (2018) · The Journal of General Physiology · doi:10.1085/jgp.201711979 · Cited by 50 · see citing works
Gormal, R. S., Padmanabhan, P., Kasula, R., Bademosi, A. T., Coakley, S., Giacomotto, J., Blum, A., Joensuu, M., Wallis, T. P., Lo, H. P., et al. (2020) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.2007443117 · Cited by 49 · see citing works
Self-citationBruzzese, A., Gil, C., Dalton, J. A. R., & Giraldo, J. (2018) · Scientific Reports · doi:10.1038/s41598-018-22735-6 · Cited by 48 · see citing works
Therapeutic Potential of Targeting ß-Arrestin
Bond, R. A., Garcia-Rojas, E. Y. L., Hegde, A., & Walker, J. K. L. (2019) · Frontiers in Pharmacology · doi:10.3389/fphar.2019.00124 · Cited by 46 · see citing works
Staus, D. P., Wingler, L. M., Pichugin, D., Prosser, R. S., & Lefkowitz, R. J. (2019) · Journal of Biological Chemistry · doi:10.1074/jbc.ac119.009848 · Cited by 46 · see citing works
Self-citationQuantitative structural assessment of graded receptor agonism
Shang, J., Brust, R., Griffin, P. R., Kamenecka, T. M., & Kojetin, D. J. (2019) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1909016116 · Cited by 45 · see citing works
β-arrestin signalling and bias in hormone-responsive GPCRs
Reiter, É., Ayoub, M. A., Pellissier, L. P., Landomiel, F., Musnier, A., Trefier, A., Gandía, J., Pascali, F. D., Tahir, S., Yvinec, R., et al. (2017) · Molecular and Cellular Endocrinology · doi:10.1016/j.mce.2017.01.052 · Cited by 44 · see citing works
A method for structure determination of GPCRs in various states
Guo, Q., He, B., Zhong, Y., Jiao, H., Ren, Y., Wang, Q., Ge, Q., Gao, Y., Liu, X., Du, Y., et al. (2023) · Nature Chemical Biology · doi:10.1038/s41589-023-01389-0 · Cited by 44 · see citing works
Gahbauer, S., Pluháčková, K., & Böckmann, R. A. (2018) · PLoS Computational Biology · doi:10.1371/journal.pcbi.1006062 · Cited by 44 · see citing works
Nanobody‐Enabled Reverse Pharmacology on G‐Protein‐Coupled Receptors
Pardon, E., Betti, C., Laeremans, T., Chevillard, F., Guillemyn, K., Kolb, P., Ballet, S., & Steyaert, J. (2018) · Angewandte Chemie International Edition · doi:10.1002/anie.201712581 · Cited by 43 · see citing works
Self-citation
- Authors
- Staus, D. P., Strachan, R. T., Manglik, A., Pani, B., Kahsai, A. W., Kim, T. H., Wingler, L. M., Ahn, S., Chatterjee, A., Masoudi, A., Kruse, A. C., Pardon, E., Steyaert, J., Weis, W. I., Prosser, R. S., Kobilka, B. K., Costa, T., & Lefkowitz, R. J.
- Affiliations
- Duke Medical Center (US)
- University of North Carolina at Chapel Hill (US)
- Stanford Medicine (US)
- University of Toronto (CA)
- Harvard University (US)
- Vrije Universiteit Brussel (BE)
- Istituto Superiore di Sanità (IT)
- Howard Hughes Medical Institute (US)
- Corresponding author
- Kobilka, Brian K.
- Costa, Tommaso
- Lefkowitz, Robert J.
- ORCID
- Venue
- Nature
- Published
- 2016-07
- Volume
- 535
- Issue
- 7612
- Pages
- 448-452
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- PMC
- PMC4961583
- OpenAlex
- W2471992026
- Cited by
- 330
- References
- 38
- Citations by year
- 2026: 11 · 2025: 18 · 2024: 23 · 2023: 28 · 2022: 26 · 2021: 46 · 2020: 36 · 2019: 47 · 2018: 43 · 2017: 39 · 2016: 13
- FWCI
- 25.13
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- OSTI OAI (U.S. Department of Energy Office of Scientific and Technical Information) (submittedVersion)
- Digital Access to Scholarship at Harvard (DASH) (Harvard University) (submittedVersion, cc-by)
- eScholarship (California Digital Library) (submittedVersion)
- VUBIR (Vrije Universiteit Brussel) (submittedVersion)
- Funders
- Howard Hughes Medical Institute
- American Heart Association
- Ministero della Salute (02351158, RF-201102351158)
- National Heart, Lung, and Blood Institute (HL 16037, R01HL070631, HL-70631, T32HL007101, R01HL016037)
- National Institute of General Medical Sciences (T32GM008294)
- National Institute of Diabetes and Digestive and Kidney Diseases (T32DK007731)
- National Institute of Neurological Disorders and Stroke (NS-028471, R01NS028471, R37NS028471)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
- Keywords
- beta2-adrenergic receptor, G protein-coupled receptor activation, receptor conformational states, three-state model, partial agonists, full agonists, ligand efficacy, receptor stabilization, orthosteric binding pocket, receptor dynamics
- MeSH terms
- Allosteric Regulation, Allosteric Site, Humans, Isoproterenol, Ligands, Models, Molecular, Protein Conformation, Receptors, Adrenergic, beta-2, Crystallography, X-Ray, Nuclear Magnetic Resonance, Biomolecular, Drug Partial Agonism, Protein Stability, Adrenergic beta-2 Receptor Agonists, Single-Domain Antibodies
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
