Structural insight into apelin receptor-G protein stoichiometry.
Journal
Nature structural & molecular biology
ISSN: 1545-9985
Titre abrégé: Nat Struct Mol Biol
Pays: United States
ID NLM: 101186374
Informations de publication
Date de publication:
07 2022
07 2022
Historique:
received:
10
03
2022
accepted:
26
05
2022
pubmed:
12
7
2022
medline:
20
7
2022
entrez:
11
7
2022
Statut:
ppublish
Résumé
The technique of cryogenic-electron microscopy (cryo-EM) has revolutionized the field of membrane protein structure and function with a focus on the dominantly observed molecular species. This report describes the structural characterization of a fully active human apelin receptor (APJR) complexed with heterotrimeric G protein observed in both 2:1 and 1:1 stoichiometric ratios. We use cryo-EM single-particle analysis to determine the structural details of both species from the same sample preparation. Protein preparations, in the presence of the endogenous peptide ligand ELA or a synthetic small molecule, both demonstrate these mixed stoichiometric states. Structural differences in G protein engagement between dimeric and monomeric APJR suggest a role for the stoichiometry of G protein-coupled receptor- (GPCR-)G protein coupling on downstream signaling and receptor pharmacology. Furthermore, a small, hydrophobic dimer interface provides a starting framework for additional class A GPCR dimerization studies. Together, these findings uncover a mechanism of versatile regulation through oligomerization by which GPCRs can modulate their signaling.
Identifiants
pubmed: 35817871
doi: 10.1038/s41594-022-00797-5
pii: 10.1038/s41594-022-00797-5
doi:
Substances chimiques
Apelin Receptors
0
Carrier Proteins
0
Receptors, G-Protein-Coupled
0
GTP-Binding Proteins
EC 3.6.1.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
688-697Subventions
Organisme : CIHR
ID : FDN-148413
Pays : Canada
Informations de copyright
© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.
Références
Sleno, R. & Hébert, T. E. Shaky ground—the nature of metastable GPCR signalling complexes. Neuropharmacology 152, 4–14 (2019).
pubmed: 30659839
doi: 10.1016/j.neuropharm.2019.01.018
Milligan, G., Ward, R. J. & Marsango, S. GPCR homo-oligomerization. Curr. Opin. Cell Biol. 57, 40–47 (2019).
pubmed: 30453145
pmcid: 7083226
doi: 10.1016/j.ceb.2018.10.007
Pétrin, D. & Hébert, T. E. The functional size of GPCRs—monomers, dimers or tetramers? Subcell Biochem. 63, 67–81 (2012).
pubmed: 23161133
doi: 10.1007/978-94-007-4765-4_4
Dupré, D. J. & Hébert, T. E. Biosynthesis and trafficking of seven transmembrane receptor signalling complexes. Cell Signal 18, 1549–1559 (2006).
pubmed: 16677801
doi: 10.1016/j.cellsig.2006.03.009
Lopez-Gimenez, J. F., Canals, M., Pediani, J. D. & Milligan, G. The alpha1b-adrenoceptor exists as a higher-order oligomer: effective oligomerization is required for receptor maturation, surface delivery, and function. Mol. Pharmacol. 71, 1015–1029 (2007).
pubmed: 17220353
doi: 10.1124/mol.106.033035
Salahpour, A. et al. Homodimerization of the beta2-adrenergic receptor as a prerequisite for cell surface targeting. J. Biol. Chem. 279, 33390–33397 (2004).
pubmed: 15155738
doi: 10.1074/jbc.M403363200
Papasergi-Scott, M. M. et al. Structures of metabotropic GABA(B) receptor. Nature 584, 310–314 (2020).
pubmed: 32580208
pmcid: 7429364
doi: 10.1038/s41586-020-2469-4
Scholler, P. et al. Allosteric nanobodies uncover a role of hippocampal mGlu2 receptor homodimers in contextual fear consolidation. Nat. Commun. 8, 1967 (2017).
pubmed: 29213077
pmcid: 5719040
doi: 10.1038/s41467-017-01489-1
Zhu, S. et al. Structure of a human synaptic GABA(A) receptor. Nature 559, 67–72 (2018).
pubmed: 29950725
pmcid: 6220708
doi: 10.1038/s41586-018-0255-3
Dijkman, P. M. et al. Dynamic tuneable G protein-coupled receptor monomer-dimer populations. Nat. Commun. 9, 1710 (2018).
pubmed: 29703992
pmcid: 5923235
doi: 10.1038/s41467-018-03727-6
Ferré, S. et al. G protein-coupled receptor oligomerization revisited: functional and pharmacological perspectives. Pharmacol Rev 66, 413–434 (2014).
pubmed: 24515647
pmcid: 3973609
doi: 10.1124/pr.113.008052
Angers, S. et al. Detection of beta 2-adrenergic receptor dimerization in living cells using bioluminescence resonance energy transfer (BRET). Proc. Natl Acad. Sci. USA 97, 3684–3689 (2000).
pubmed: 10725388
pmcid: 16300
Calebiro, D. et al. Single-molecule analysis of fluorescently labeled G-protein-coupled receptors reveals complexes with distinct dynamics and organization. Proc. Natl Acad. Sci. USA 110, 743–748 (2013).
pubmed: 23267088
doi: 10.1073/pnas.1205798110
Möller, J. et al. Single-molecule analysis reveals agonist-specific dimer formation of µ-opioid receptors. Nat. Chem. Biol. 16, 946–954 (2020).
pubmed: 32541966
doi: 10.1038/s41589-020-0566-1
Fiorentini, C., Busi, C., Spano, P. & Missale, C. Dimerization of dopamine D1 and D3 receptors in the regulation of striatal function. Curr. Opin. Pharmacol. 10, 87–92 (2010).
pubmed: 19837631
doi: 10.1016/j.coph.2009.09.008
Trettel, F. et al. Ligand-independent CXCR2 dimerization. J. Biol. Chem. 278, 40980–40988 (2003).
pubmed: 12888558
doi: 10.1074/jbc.M306815200
Işbilir, A. et al. Advanced fluorescence microscopy reveals disruption of dynamic CXCR4 dimerization by subpocket-specific inverse agonists. Proc. Natl Acad. Sci. USA 117, 29144–29154 (2020).
pubmed: 33148803
pmcid: 7682396
doi: 10.1073/pnas.2013319117
Herrick-Davis, K., Grinde, E., Harrigan, T. J. & Mazurkiewicz, J. E. Inhibition of serotonin 5-hydroxytryptamine2c receptor function through heterodimerization: receptor dimers bind two molecules of ligand and one G-protein. J. Biol. Chem. 280, 40144–40151 (2005).
pubmed: 16195233
doi: 10.1074/jbc.M507396200
Zhao, D. Y. et al. Cryo-EM structure of the native rhodopsin dimer in nanodiscs. J. Biol. Chem. 294, 14215–14230 (2019).
pubmed: 31399513
pmcid: 6768649
doi: 10.1074/jbc.RA119.010089
Liang, Y. et al. Organization of the G protein-coupled receptors rhodopsin and opsin in native membranes. J. Biol. Chem. 278, 21655–21662 (2003).
pubmed: 12663652
doi: 10.1074/jbc.M302536200
Huang, J., Chen, S., Zhang, J. J. & Huang, X. Y. Crystal structure of oligomeric β1-adrenergic G protein-coupled receptors in ligand-free basal state. Nat. Struct. Mol. Biol. 20, 419–425 (2013).
pubmed: 23435379
pmcid: 3618578
doi: 10.1038/nsmb.2504
Manglik, A. et al. Crystal structure of the µ-opioid receptor bound to a morphinan antagonist. Nature 485, 321–326 (2012).
pubmed: 22437502
pmcid: 3523197
doi: 10.1038/nature10954
Wu, B. et al. Structures of the CXCR4 chemokine GPCR with small-molecule and cyclic peptide antagonists. Science 330, 1066–1071 (2010).
pubmed: 20929726
pmcid: 3074590
doi: 10.1126/science.1194396
Siddiquee, K., Hampton, J., McAnally, D., May, L. & Smith, L. The apelin receptor inhibits the angiotensin II type 1 receptor via allosteric trans-inhibition. Br. J. Pharmacol. 168, 1104–1117 (2013).
pubmed: 22935142
pmcid: 3594671
doi: 10.1111/j.1476-5381.2012.02192.x
Cai, X., Bai, B., Zhang, R., Wang, C. & Chen, J. Apelin receptor homodimer-oligomers revealed by single-molecule imaging and novel G protein-dependent signaling. Sci. Rep. 7, 40335 (2017).
pubmed: 28091541
pmcid: 5238433
doi: 10.1038/srep40335
Chun, H. J. et al. Apelin signaling antagonizes Ang II effects in mouse models of atherosclerosis. J. Clin. Invest. 118, 3343–3354 (2008).
pubmed: 18769630
pmcid: 2525695
Li, Y. et al. Heterodimerization of human apelin and kappa opioid receptors: roles in signal transduction. Cell Signal 24, 991–1001 (2012).
pubmed: 22200678
doi: 10.1016/j.cellsig.2011.12.012
Ji, B. et al. Roles for heterodimerization of APJ and B2R in promoting cell proliferation via ERK1/2-eNOS signaling pathway. Cell Signal 73, 109671 (2020).
pubmed: 32407761
doi: 10.1016/j.cellsig.2020.109671
O’Dowd, B. F. et al. A human gene that shows identity with the gene encoding the angiotensin receptor is located on chromosome 11. Gene 136, 355–360 (1993).
pubmed: 8294032
doi: 10.1016/0378-1119(93)90495-O
Edinger, A. L. et al. An orphan seven-transmembrane domain receptor expressed widely in the brain functions as a coreceptor for human immunodeficiency virus type 1 and simian immunodeficiency virus. J. Virol. 72, 7934–7940 (1998).
pubmed: 9733831
pmcid: 110125
doi: 10.1128/JVI.72.10.7934-7940.1998
Chng, S. C., Ho, L., Tian, J. & Reversade, B. Elabela: a hormone essential for heart development signals via the apelin receptor. Dev. Cell 27, 672–680 (2013).
pubmed: 24316148
doi: 10.1016/j.devcel.2013.11.002
Cox, C. M., D’Agostino, S. L., Miller, M. K., Heimark, R. L. & Krieg, P. A. Apelin, the ligand for the endothelial G-protein-coupled receptor, APJ, is a potent angiogenic factor required for normal vascular development of the frog embryo. Dev. Biol. 296, 177–189 (2006).
pubmed: 16750822
doi: 10.1016/j.ydbio.2006.04.452
Kasai, A. et al. Retardation of retinal vascular development in apelin-deficient mice. Arterioscler. Thromb. Vasc. Biol. 28, 1717–1722 (2008).
pubmed: 18599802
doi: 10.1161/ATVBAHA.108.163402
Wang, Z. et al. Elabela-apelin receptor signaling pathway is functional in mammalian systems. Sci. Rep. 5, 8170 (2015).
pubmed: 25639753
pmcid: 4313117
doi: 10.1038/srep08170
Marsault, E. et al. The apelinergic system: a perspective on challenges and opportunities in cardiovascular and metabolic disorders. Ann. N. Y. Acad. Sci. 1455, 12–33 (2019).
pubmed: 31236974
pmcid: 6834863
doi: 10.1111/nyas.14123
Scimia, M. C. et al. APJ acts as a dual receptor in cardiac hypertrophy. Nature 488, 394–398 (2012).
pubmed: 22810587
pmcid: 3422434
doi: 10.1038/nature11263
Ma, Y. et al. Structural basis for apelin control of the human apelin receptor. Structure 25, 858–866 (2017).
pubmed: 28528775
doi: 10.1016/j.str.2017.04.008
Pauli, A. et al. Toddler: an embryonic signal that promotes cell movement via Apelin receptors. Science 343, 1248636 (2014).
pubmed: 24407481
pmcid: 4107353
doi: 10.1126/science.1248636
Perjés, Á. et al. Characterization of apela, a novel endogenous ligand of apelin receptor, in the adult heart. Basic Res. Cardiol. 111, 2 (2016).
pubmed: 26611206
doi: 10.1007/s00395-015-0521-6
Dagamajalu, S. et al. The network map of Elabela signaling pathway in physiological and pathological conditions. J. Cell Commun. Signal. 16, 145–154 (2021).
pubmed: 34339006
pmcid: 8688647
doi: 10.1007/s12079-021-00640-4
Sato, T. et al. ELABELA-APJ axis protects from pressure overload heart failure and angiotensin II-induced cardiac damage. Cardiovasc. Res. 113, 760–769 (2017).
pubmed: 28371822
doi: 10.1093/cvr/cvx061
Deng, C., Chen, H., Yang, N., Feng, Y. & Hsueh, A. J. Apela regulates fluid homeostasis by binding to the APJ receptor to activate Gi signaling. J. Biol. Chem. 290, 18261–18268 (2015).
pubmed: 25995451
pmcid: 4513087
doi: 10.1074/jbc.M115.648238
Ho, L. et al. ELABELA deficiency promotes preeclampsia and cardiovascular malformations in mice. Science 357, 707–713 (2017).
pubmed: 28663440
doi: 10.1126/science.aam6607
Chapman, F. A. et al. The therapeutic potential of apelin in kidney disease. Nat. Rev. Nephrol. 17, 840–853 (2021).
pubmed: 34389827
pmcid: 8361827
doi: 10.1038/s41581-021-00461-z
Chen, H. et al. ELABELA and an ELABELA fragment protect against AKI. J. Am. Soc. Nephrol. 28, 2694–2707 (2017).
pubmed: 28583915
pmcid: 5576937
doi: 10.1681/ASN.2016111210
Liet, B., Nys, N. & Siegfried, G. Elabela/toddler: new peptide with a promising future in cancer diagnostic and therapy. Biochim. Biophys. Acta, Mol. Cell. Res. 1868, 119065 (2021).
doi: 10.1016/j.bbamcr.2021.119065
Chen, N. et al. Triazole agonists of the APJ receptor. Patent WO2016187308A1 (2016).
Ason, B. et al. Cardiovascular response to small-molecule APJ activation. JCI Insight 5, e132898 (2020).
pmcid: 7205427
doi: 10.1172/jci.insight.132898
Maeda, S. et al. Development of an antibody fragment that stabilizes GPCR/G-protein complexes. Nat. Commun. 9, 3712 (2018).
pubmed: 30213947
pmcid: 6137068
doi: 10.1038/s41467-018-06002-w
Shen, C. et al. Structural basis of GABAB receptor–Gi protein coupling. Nature 594, 594–598 (2021).
pubmed: 33911284
pmcid: 8222003
doi: 10.1038/s41586-021-03507-1
Lin, S. et al. Structures of G(i)-bound metabotropic glutamate receptors mGlu2 and mGlu4. Nature 594, 583–588 (2021).
pubmed: 34135510
doi: 10.1038/s41586-021-03495-2
Velazhahan, V. et al. Structure of the class D GPCR Ste2 dimer coupled to two G proteins. Nature 589, 148–153 (2021).
pubmed: 33268889
doi: 10.1038/s41586-020-2994-1
Ballesteros, J. A. & Weinstein, H. In Methods in Neurosciences Vol. 25 (ed Sealfon, S. C.) 366–428 (Academic Press, 1995).
Pettersen, E. F. et al. UCSF Chimera–a visualization system for exploratory research and analysis. J. Comput. Chem. 25, 1605–1612 (2004).
pubmed: 15264254
doi: 10.1002/jcc.20084
Reggio, P. H. Computational methods in drug design: modeling G protein-coupled receptor monomers, dimers, and oligomers. AAAPS. J. 8, E322–E336 (2006).
doi: 10.1007/BF02854903
Terrillon, S. et al. Oxytocin and vasopressin V1a and V2 receptors form constitutive homo- and heterodimers during biosynthesis. Mol. Endocrinol. 17, 677–691 (2003).
pubmed: 12554793
doi: 10.1210/me.2002-0222
Kroeze, W. K. et al. PRESTO-Tango as an open-source resource for interrogation of the druggable human GPCRome. Nat. Struct. Mol. Biol. 22, 362–369 (2015).
pubmed: 25895059
pmcid: 4424118
doi: 10.1038/nsmb.3014
Barnea, G. et al. The genetic design of signaling cascades to record receptor activation. Proc. Natl Acad. Sci. USA 105, 64–69 (2008).
pubmed: 18165312
doi: 10.1073/pnas.0710487105
Ma, Y. et al. Structure-guided discovery of a single-domain antibody agonist against human apelin receptor. Sci. Adv. 6, eaax7379 (2020).
pubmed: 31998837
pmcid: 6962038
doi: 10.1126/sciadv.aax7379
Zhang, H. et al. Structural basis for selectivity and diversity in angiotensin II receptors. Nature 544, 327–332 (2017).
pubmed: 28379944
pmcid: 5525545
doi: 10.1038/nature22035
Scott, I. C. et al. The G protein-coupled receptor agtrl1b regulates early development of myocardial progenitors. Dev. Cell 12, 403–413 (2007).
pubmed: 17336906
doi: 10.1016/j.devcel.2007.01.012
Murza, A. et al. Discovery and structure–activity relationship of a bioactive fragment of ELABELA that modulates vascular and cardiac functions. J. Med. Chem. 59, 2962–2972 (2016).
pubmed: 26986036
doi: 10.1021/acs.jmedchem.5b01549
Read, C. et al. International Union of Basic and Clinical Pharmacology. CVII. Structure and pharmacology of the apelin receptor with a recommendation that ELABELA/toddler is a second endogenous peptide ligand. Pharmacol. Rev. 71, 467–502 (2019).
pubmed: 31492821
pmcid: 6731456
doi: 10.1124/pr.119.017533
Trân, K. et al. Structure–activity relationship and bioactivity of short analogues of ELABELA as agonists of the apelin receptor. J. Med. Chem. 64, 602–615 (2021).
pubmed: 33350824
doi: 10.1021/acs.jmedchem.0c01547
Felce, J. H., Davis, S. J. & Klenerman, D. Single-molecule analysis of G protein-coupled receptor stoichiometry: approaches and limitations. Trends Pharmacol. Sci. 39, 96–108 (2018).
pubmed: 29122289
doi: 10.1016/j.tips.2017.10.005
Murata, K. & Wolf, M. Cryo-electron microscopy for structural analysis of dynamic biological macromolecules. Biochim. Biophys. Acta, Gen. Subj. 1862, 324–334 (2018).
doi: 10.1016/j.bbagen.2017.07.020
Lau, C., Hunter, M. J., Stewart, A., Perozo, E. & Vandenberg, J. I. Never at rest: insights into the conformational dynamics of ion channels from cryo-electron microscopy. J. Physiol. 596, 1107–1119 (2018).
pubmed: 29377132
pmcid: 5878226
doi: 10.1113/JP274888
Punjani, A., Rubinstein, J. L., Fleet, D. J. & Brubaker, M. A. cryoSPARC: algorithms for rapid unsupervised cryo-EM structure determination. Nat. Methods 14, 290–296 (2017).
pubmed: 28165473
doi: 10.1038/nmeth.4169
Velazhahan, V. et al. Structure of the class D GPCR Ste2 dimer coupled to two G proteins. Nature 589, 148–153 (2020).
pubmed: 33268889
pmcid: 7116888
doi: 10.1038/s41586-020-2994-1
Koehl, A. et al. Structural insights into the activation of metabotropic glutamate receptors. Nature 566, 79–84 (2019).
pubmed: 30675062
pmcid: 6709600
doi: 10.1038/s41586-019-0881-4
Mao, C. et al. Cryo-EM structures of inactive and active GABA(B) receptor. Cell Res. 30, 564–573 (2020).
pubmed: 32494023
pmcid: 7343782
doi: 10.1038/s41422-020-0350-5
Chun, E. et al. Fusion partner toolchest for the stabilization and crystallization of G protein-coupled receptors. Structure 20, 967–976 (2012).
pubmed: 22681902
pmcid: 3375611
doi: 10.1016/j.str.2012.04.010
Hua, T. et al. Activation and signaling mechanism revealed by cannabinoid receptor-G(i) complex structures. Cell 180, 655–665 (2020).
pubmed: 32004463
pmcid: 7898353
doi: 10.1016/j.cell.2020.01.008
Bepler, T. et al. Positive-unlabeled convolutional neural networks for particle picking in cryo-electron micrographs. Nat. Methods 16, 1153–1160 (2019).
pubmed: 31591578
pmcid: 6858545
doi: 10.1038/s41592-019-0575-8
Sanchez-Garcia, R. et al. DeepEMhancer: a deep learning solution for cryo-EM volume post-processing. Commun. Biol. 4, 874 (2021).
pubmed: 34267316
pmcid: 8282847
doi: 10.1038/s42003-021-02399-1
Emsley, P., Lohkamp, B., Scott, W. G. & Cowtan, K. Features and development of Coot. Acta Crystallogr. D. Biol. Crystallogr. 66, 486–501 (2010).
pubmed: 20383002
pmcid: 2852313
doi: 10.1107/S0907444910007493
McCoy, A. J. et al. Phaser crystallographic software. J. Appl. Crystallogr. 40, 658–674 (2007).
pubmed: 19461840
pmcid: 2483472
doi: 10.1107/S0021889807021206
Chen, V. B. et al. MolProbity: all-atom structure validation for macromolecular crystallography. Acta Crystallogr. D. Biol. Crystallogr. 66, 12–21 (2010).
pubmed: 20057044
doi: 10.1107/S0907444909042073
Caffrey, M. & Cherezov, V. Crystallizing membrane proteins using lipidic mesophases. Nat. Protoc. 4, 706–731 (2009).
pubmed: 19390528
pmcid: 2732203
doi: 10.1038/nprot.2009.31
Kabsch, W. Integration, scaling, space-group assignment and post-refinement. Acta Crystallogr. D. Biol. Crystallogr. 66, 133–144 (2010).
pubmed: 20124693
pmcid: 2815666
doi: 10.1107/S0907444909047374
Murshudov, G. N., Vagin, A. A. & Dodson, E. J. Refinement of macromolecular structures by the maximum-likelihood method. Acta Crystallogr. D. Biol. Crystallogr. 53, 240–255 (1997).
pubmed: 15299926
doi: 10.1107/S0907444996012255
Smart, K. M., Blake, C., Staines, A., Thacker, M. & Doody, C. Mechanisms-based classifications of musculoskeletal pain: part 1 of 3: symptoms and signs of central sensitisation in patients with low back (± leg) pain. Man. Ther. 17, 336–344 (2012).
pubmed: 22534654
doi: 10.1016/j.math.2012.03.013
Ritchie, T. K. et al. Chapter 11—reconstitution of membrane proteins in phospholipid bilayer nanodiscs. Methods Enzymol. 464, 211–231 (2009).
pubmed: 19903557
pmcid: 4196316
doi: 10.1016/S0076-6879(09)64011-8
Olsen, R. H. J. et al. TRUPATH, an open-source biosensor platform for interrogating the GPCR transducerome. Nat. Chem. Biol. 16, 841–849 (2020).
pubmed: 32367019
pmcid: 7648517
doi: 10.1038/s41589-020-0535-8