{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T02:44:02Z","timestamp":1785811442436,"version":"3.56.0"},"reference-count":41,"publisher":"Springer Science and Business Media LLC","license":[{"start":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T00:00:00Z","timestamp":1785801600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T00:00:00Z","timestamp":1785801600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Naunyn-Schmiedeberg's Arch Pharmacol"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The bradykinin 2 receptor (B2R) is one of two members of the kinin receptor family and is a G protein\u2013coupled receptor (GPCR) that regulates important physiological processes including pain, inflammation, and cardiovascular homeostasis. Functionally, B2R is activated by the kinin peptides, bradykinin and kallidin. Carboxypeptidase cleavage of bradykinin and kallidin results in the formation of des-Arg9-bradykinin (DABK) and des-Arg10-kallidin (DAKD) respectively, which are classically described as agonists of the bradykinin 1 receptor (B1R), the other member of the kinin family. Affinity binding studies have reported that DABK and DAKD can also bind to B2R. Upon activation, B2R signals through the recruitment of heterotrimeric G proteins, and earlier work has highlighted the promiscuous G protein coupling to B2R in response to bradykinin. However, a comprehensive G protein activation profile of the B2R receptor when activated by the other endogenous agonists has not yet been established. In this study, we used Bioluminescence Resonance Energy Transfer (BRET)\u2013based in vitro assays to monitor the coupling of 14 different G\u03b1 proteins upon B2R activation. Our results show that there is a shift in G protein activation profiles and kinetics among the different peptides. By analyzing responses across a range of agonist concentrations, we further identified biased signaling among these peptides, with differential activation of specific G\u03b1 subtypes. These results improve understanding of intracellular mechanisms through which the kinin system regulates different aspects of its physiological role and provide valuable information for the design of drugs targeting the kinin receptors.<\/jats:p>","DOI":"10.1007\/s00210-026-05758-z","type":"journal-article","created":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T01:47:48Z","timestamp":1785808068000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["G protein signaling capabilities at the bradykinin 2 receptor"],"prefix":"10.1007","author":[{"given":"Shradha V.","family":"Darira","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laurie P.","family":"Sutton","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,8,4]]},"reference":[{"key":"5758_CR1","doi-asserted-by":"publisher","first-page":"230","DOI":"10.1124\/molpharm.123.000722","volume":"104","author":"R Alabdali","year":"2023","unstructured":"Alabdali R, Franchini L, Orlandi C (2023a) G\u03b1 protein signaling bias at serotonin 1A receptor. Mol Pharmacol 104:230\u2013238","journal-title":"Mol Pharmacol"},{"key":"5758_CR2","doi-asserted-by":"publisher","first-page":"230","DOI":"10.1124\/molpharm.123.000722","volume":"104","author":"R Alabdali","year":"2023","unstructured":"Alabdali R, Franchini L, Orlandi C (2023b) Galpha protein signaling bias at serotonin 1A receptor. Mol Pharmacol 104:230\u2013238","journal-title":"Mol Pharmacol"},{"key":"5758_CR3","doi-asserted-by":"crossref","unstructured":"Black JW, Leff P, Shankley NP, Wood J (2010) An operational model of pharmacological agonism: the effect of E\/[A] curve shape on agonist dissociation constant estimation. 1985. British journal of pharmacology 160 Suppl 1: S54\u201364","DOI":"10.1111\/j.1476-5381.2010.00855.x"},{"key":"5758_CR4","doi-asserted-by":"publisher","first-page":"789","DOI":"10.1038\/nrd.2018.135","volume":"17","author":"AP Campbell","year":"2018","unstructured":"Campbell AP, Smrcka AV (2018) Targeting G protein-coupled receptor signalling by blocking G proteins. Nat Rev Drug Discov 17:789\u2013803","journal-title":"Nat Rev Drug Discov"},{"key":"5758_CR5","doi-asserted-by":"publisher","DOI":"10.1126\/scisignal.aav0320","author":"T Che","year":"2021","unstructured":"Che T, Dwivedi-Agnihotri H, Shukla AK, Roth BL (2021) Biased ligands at opioid receptors: current status and future directions. Sci Signal. https:\/\/doi.org\/10.1126\/scisignal.aav0320","journal-title":"Sci Signal"},{"key":"5758_CR6","doi-asserted-by":"publisher","first-page":"2305","DOI":"10.1161\/01.CIR.100.23.2305","volume":"100","author":"LJ Dell\u2019Italia","year":"1999","unstructured":"Dell\u2019Italia LJ, Oparil S (1999) Bradykinin in the heart. Circulation 100:2305\u20132307","journal-title":"Circulation"},{"key":"5758_CR7","doi-asserted-by":"publisher","first-page":"73","DOI":"10.1021\/acsptsci.8b00026","volume":"1","author":"A Glukhova","year":"2018","unstructured":"Glukhova A, Draper-Joyce CJ, Sunahara RK, Christopoulos A, Wootten D, Sexton PM (2018) Rules of engagement: GPCRs and G proteins. ACS Pharmacol Transl Sci 1:73\u201383","journal-title":"ACS Pharmacol Transl Sci"},{"key":"5758_CR8","doi-asserted-by":"publisher","first-page":"17901","DOI":"10.1074\/jbc.274.25.17901","volume":"274","author":"A Gohla","year":"1999","unstructured":"Gohla A, Offermanns S, Wilkie TM, Schultz G (1999) Differential involvement of Galpha12 and Galpha13 in receptor-mediated stress fiber formation. J Biol Chem 274:17901\u201317907","journal-title":"J Biol Chem"},{"key":"5758_CR9","doi-asserted-by":"publisher","first-page":"11820","DOI":"10.1073\/pnas.2000712117","volume":"117","author":"I Gomes","year":"2020","unstructured":"Gomes I, Sierra S, Lueptow L, Gupta A, Gouty S, Margolis EB, Cox BM, Devi LA (2020) Biased signaling by endogenous opioid peptides. Proc Natl Acad Sci U S A 117:11820\u201311828","journal-title":"Proc Natl Acad Sci U S A"},{"key":"5758_CR10","doi-asserted-by":"publisher","first-page":"2376","DOI":"10.1021\/acs.biochem.5c00022","volume":"64","author":"GM Grieble","year":"2025","unstructured":"Grieble GM, Knapp BI, Bidlack JM (2025) Mu opioid receptor positive allosteric modulator BMS-986122 confers agonist-dependent G protein subtype signaling bias. Biochemistry 64:2376\u20132393","journal-title":"Biochemistry"},{"issue":"1","key":"5758_CR11","doi-asserted-by":"publisher","first-page":"341","DOI":"10.1038\/s41467-017-02661-3","volume":"9","author":"M Grundmann","year":"2018","unstructured":"Grundmann M, Merten N, Malfacini D, Inoue A, Preis P, Simon K, Ruttiger N, Ziegler N, Benkel T, Schmitt NK, Ishida S, Muller I, Reher R, Kawakami K, Inoue A, Rick U, Kuhl T, Imhof D, Aoki J, Konig GM, Hoffmann C, Gomeza J, Wess J, Kostenis E (2018) Lack of beta-arrestin signaling in the absence of active G proteins. Nat Commun 9:341","journal-title":"Nat Commun"},{"key":"5758_CR12","doi-asserted-by":"publisher","first-page":"1015","DOI":"10.1016\/j.cellsig.2009.02.017","volume":"21","author":"B Hollins","year":"2009","unstructured":"Hollins B, Kuravi S, Digby GJ, Lambert NA (2009) The c-terminus of GRK3 indicates rapid dissociation of G protein heterotrimers. Cell Signal 21:1015\u20131021","journal-title":"Cell Signal"},{"key":"5758_CR13","doi-asserted-by":"publisher","DOI":"10.1111\/jnc.16248","volume":"169","author":"M Holtje","year":"2025","unstructured":"Holtje M, Wolkowicz A, Brunk I, Baron J, Ahnert-Hilger G (2025) Galpha(o1) and Galpha(o1)\/Galpha(o2) deletion differentially affect hippocampal mossy fiber tract anatomy and neuronal morphogenesis. J Neurochem 169:e16248","journal-title":"J Neurochem"},{"key":"5758_CR14","doi-asserted-by":"publisher","first-page":"284","DOI":"10.1038\/nchembio.2551","volume":"14","author":"L Joedicke","year":"2018","unstructured":"Joedicke L, Mao J, Kuenze G, Reinhart C, Kalavacherla T, Jonker HRA, Richter C, Schwalbe H, Meiler J, Preu J, Michel H, Glaubitz C (2018) The molecular basis of subtype selectivity of human kinin G-protein-coupled receptors. Nat Chem Biol 14:284\u2013290","journal-title":"Nat Chem Biol"},{"key":"5758_CR15","doi-asserted-by":"publisher","first-page":"12543","DOI":"10.1007\/s00210-025-03999-y","volume":"398","author":"N Katnahji","year":"2025","unstructured":"Katnahji N, Matthes J (2025) Opposite effects of Galpha(i2) or Galpha(i3) deficiency on reduced basal density and attenuated beta-adrenergic response of ventricular Ca(2+) currents in myocytes of mice overexpressing the cardiac beta(1)-adrenoceptor. Naunyn Schmiedebergs Arch Pharmacol 398:12543\u201312549","journal-title":"Naunyn Schmiedebergs Arch Pharmacol"},{"key":"5758_CR16","doi-asserted-by":"publisher","first-page":"205","DOI":"10.1038\/nrd3954","volume":"12","author":"T Kenakin","year":"2013","unstructured":"Kenakin T, Christopoulos A (2013) Signalling bias in new drug discovery: detection, quantification and therapeutic impact. Nat Rev Drug Discov 12:205\u2013216","journal-title":"Nat Rev Drug Discov"},{"key":"5758_CR17","doi-asserted-by":"publisher","first-page":"193","DOI":"10.1021\/cn200111m","volume":"3","author":"T Kenakin","year":"2012","unstructured":"Kenakin T, Watson C, Muniz-Medina V, Christopoulos A, Novick S (2012) A simple method for quantifying functional selectivity and agonist bias. ACS Chem Neurosci 3:193\u2013203","journal-title":"ACS Chem Neurosci"},{"key":"5758_CR18","doi-asserted-by":"publisher","first-page":"3651","DOI":"10.1111\/bph.15811","volume":"179","author":"P Kolb","year":"2022","unstructured":"Kolb P, Kenakin T, Alexander SPH, Bermudez M, Bohn LM, Breinholt CS, Bouvier M, Hill SJ, Kostenis E, Martemyanov KA, Neubig RR, Onaran HO, Rajagopal S, Roth BL, Selent J, Shukla AK, Sommer ME, Gloriam DE (2022) Community guidelines for GPCR ligand bias: IUPHAR review 32. Br J Pharmacol 179:3651\u20133674","journal-title":"Br J Pharmacol"},{"key":"5758_CR19","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1124\/pr.57.1.2","volume":"57","author":"LM Leeb-Lundberg","year":"2005","unstructured":"Leeb-Lundberg LM, Marceau F, M\u00fcller-Esterl W, Pettibone DJ, Zuraw BL (2005) International union of pharmacology. XLV. Classification of the kinin receptor family: from molecular mechanisms to pathophysiological consequences. Pharmacol Rev 57:27\u201377","journal-title":"Pharmacol Rev"},{"key":"5758_CR20","doi-asserted-by":"publisher","first-page":"2168","DOI":"10.1172\/JCI116818","volume":"92","author":"JK Liao","year":"1993","unstructured":"Liao JK, Homcy CJ (1993) The G proteins of the G alpha i and G alpha q family couple the bradykinin receptor to the release of endothelium-derived relaxing factor. J Clin Invest 92:2168\u20132172","journal-title":"J Clin Invest"},{"issue":"Pt 1","key":"5758_CR21","doi-asserted-by":"publisher","first-page":"109","DOI":"10.1042\/bj3130109","volume":"313","author":"C Liebmann","year":"1996","unstructured":"Liebmann C, Graness A, Ludwig B, Adomeit A, Boehmer A, Boehmer FD, N\u00fcrnberg B, Wetzker R (1996) Dual bradykinin B2 receptor signalling in A431 human epidermoid carcinoma cells: activation of protein kinase C is counteracted by a GS-mediated stimulation of the cyclic AMP pathway. Biochem J 313(Pt 1):109\u2013118","journal-title":"Biochem J"},{"key":"5758_CR22","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1124\/molpharm.122.000487","volume":"102","author":"SM Mack","year":"2022","unstructured":"Mack SM, Gomes I, Fakira AK, Duarte ML, Gupta A, Fricker L, Devi LA (2022) GPR83 engages endogenous peptides from two distinct precursors to elicit differential signaling. Mol Pharmacol 102:29\u201338","journal-title":"Mol Pharmacol"},{"key":"5758_CR23","doi-asserted-by":"publisher","first-page":"845","DOI":"10.1038\/nrd1522","volume":"3","author":"F Marceau","year":"2004","unstructured":"Marceau F, Regoli D (2004) Bradykinin receptor ligands: therapeutic perspectives. Nat Rev Drug Discov 3:845\u2013852","journal-title":"Nat Rev Drug Discov"},{"key":"5758_CR24","doi-asserted-by":"publisher","first-page":"107","DOI":"10.1007\/978-1-4939-2914-6_8","volume":"1335","author":"I Masuho","year":"2015","unstructured":"Masuho I, Martemyanov KA, Lambert NA (2015a) Monitoring G protein activation in cells with BRET. Methods Mol Biol 1335:107\u2013113","journal-title":"Methods Mol Biol"},{"issue":"405","key":"5758_CR25","doi-asserted-by":"publisher","first-page":"ra123","DOI":"10.1126\/scisignal.aab4068","volume":"8","author":"I Masuho","year":"2015","unstructured":"Masuho I, Ostrovskaya O, Kramer GM, Jones CD, Xie K, Martemyanov KA (2015b) Distinct profiles of functional discrimination among G proteins determine the actions of G protein-coupled receptors. Sci Signal 8:ra123","journal-title":"Sci Signal"},{"key":"5758_CR26","doi-asserted-by":"publisher","first-page":"104","DOI":"10.1124\/molpharm.123.000795","volume":"105","author":"TW McCullock","year":"2024","unstructured":"McCullock TW, Cardani LP, Kammermeier PJ (2024) Signaling specificity and kinetics of the human metabotropic glutamate receptors. Mol Pharmacol 105:104\u2013115","journal-title":"Mol Pharmacol"},{"key":"5758_CR27","doi-asserted-by":"crossref","unstructured":"M\u00f6nnich D, Humphrys L, Hoering C, Hoare B, Forster L, Pockes S (2024) Activation of multiple G protein pathways to characterize the five dopamine receptor subtypes using bioluminescence technology. ACS Pharmacology & Translational Science 7","DOI":"10.1021\/acsptsci.3c00339"},{"key":"5758_CR28","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2021.108718","volume":"34","author":"BS Muntean","year":"2021","unstructured":"Muntean BS, Masuho I, Dao M, Sutton LP, Zucca S, Iwamoto H, Patil DN, Wang D, Birnbaumer L, Blakely RD, Grill B, Martemyanov KA (2021) Galphao is a major determinant of cAMP signaling in the pathophysiology of movement disorders. Cell Rep 34:108718","journal-title":"Cell Rep"},{"key":"5758_CR29","doi-asserted-by":"publisher","first-page":"489","DOI":"10.1038\/s41401-022-00982-8","volume":"44","author":"JK Shen","year":"2023","unstructured":"Shen JK, Zhang HT (2023) Function and structure of bradykinin receptor 2 for drug discovery. Acta Pharmacol Sin 44:489\u2013498","journal-title":"Acta Pharmacol Sin"},{"key":"5758_CR30","doi-asserted-by":"publisher","first-page":"20845","DOI":"10.1074\/jbc.M110.173237","volume":"286","author":"B Sivertsen","year":"2011","unstructured":"Sivertsen B, Lang M, Frimurer TM, Holliday ND, Bach A, Els S, Engelstoft MS, Petersen PS, Madsen AN, Schwartz TW, Beck-Sickinger AG, Holst B (2011) Unique interaction pattern for a functionally biased ghrelin receptor agonist. J Biol Chem 286:20845\u201320860","journal-title":"J Biol Chem"},{"key":"5758_CR31","doi-asserted-by":"publisher","DOI":"10.1124\/molpharm.123.000818","author":"JS Smith","year":"2024","unstructured":"Smith JS, Hilibrand AS, Skiba MA, Dates AN, Calvillo-Miranda VG, Kruse AC (2024) <strong>The M3 muscarinic acetylcholine receptor can signal through multiple G protein families<\/strong>. Mol Pharmacol. https:\/\/doi.org\/10.1124\/molpharm.123.000818","journal-title":"Mol Pharmacol"},{"key":"5758_CR32","doi-asserted-by":"publisher","first-page":"251","DOI":"10.1124\/mol.117.111062","volume":"93","author":"K Sriram","year":"2018","unstructured":"Sriram K, Insel PA (2018) G protein-coupled receptors as targets for approved drugs: how many targets and how many drugs? Mol Pharmacol 93:251\u2013258","journal-title":"Mol Pharmacol"},{"key":"5758_CR33","doi-asserted-by":"publisher","first-page":"7726","DOI":"10.1074\/jbc.RA119.011984","volume":"295","author":"E van den Bos","year":"2020","unstructured":"van den Bos E, Ambrosy B, Horsthemke M, Walbaum S, Bachg AC, Wettschureck N, Innamorati G, Wilkie TM, Hanley PJ (2020) Knockout mouse models reveal the contributions of G protein subunits to complement C5a receptor-mediated chemotaxis. J Biol Chem 295:7726\u20137742","journal-title":"J Biol Chem"},{"key":"5758_CR34","doi-asserted-by":"publisher","first-page":"226","DOI":"10.1016\/j.chembiol.2021.07.004","volume":"29","author":"E Von Moo","year":"2022","unstructured":"Von Moo E, Harps\u00f8e K, Hauser AS, Masuho I, Br\u00e4uner-Osborne H, Gloriam DE, Martemyanov KA (2022) Ligand-directed bias of G protein signaling at the dopamine D(2) receptor. Cell Chem Biol 29:226-238.e224","journal-title":"Cell Chem Biol"},{"key":"5758_CR35","doi-asserted-by":"publisher","first-page":"373","DOI":"10.1021\/acsptsci.2c00020","volume":"5","author":"JH Voss","year":"2022","unstructured":"Voss JH, Mahardhika AB, Inoue A, M\u00fcller CE (2022) Agonist-dependent coupling of the promiscuous adenosine A2B receptor to G\u03b1 protein subunits. ACS Pharmacol Transl Sci 5:373\u2013386","journal-title":"ACS Pharmacol Transl Sci"},{"key":"5758_CR36","doi-asserted-by":"publisher","first-page":"1159","DOI":"10.1152\/physrev.00003.2005","volume":"85","author":"N Wettschureck","year":"2005","unstructured":"Wettschureck N, Offermanns S (2005) Mammalian G proteins and their cell type specific functions. Physiol Rev 85:1159\u20131204","journal-title":"Physiol Rev"},{"key":"5758_CR37","doi-asserted-by":"publisher","first-page":"54","DOI":"10.1186\/1744-8069-9-54","volume":"9","author":"LN Wirotanseng","year":"2013","unstructured":"Wirotanseng LN, Kuner R, Tappe-Theodor A (2013) Gq rather than G11 preferentially mediates nociceptor sensitization. Mol Pain 9:54","journal-title":"Mol Pain"},{"key":"5758_CR38","doi-asserted-by":"publisher","first-page":"24907","DOI":"10.1074\/jbc.M001051200","volume":"275","author":"P Xie","year":"2000","unstructured":"Xie P, Browning DD, Hay N, Mackman N, Ye RD (2000) Activation of NF-kappa B by bradykinin through a Galpha(q)- and Gbeta gamma-dependent pathway that involves phosphoinositide 3-kinase and Akt. J Biol Chem 275:24907\u201324914","journal-title":"J Biol Chem"},{"key":"5758_CR39","doi-asserted-by":"publisher","first-page":"227","DOI":"10.1006\/mcbr.1999.0136","volume":"1","author":"X Yang","year":"1999","unstructured":"Yang X, Taylor L, Polgar P (1999) Effect of the G-protein, G alpha(i2), and G alpha(i3) subunit knockdown on bradykinin-induced signal transduction in rat-1 cells. Mol Cell Biol Res Commun 1:227\u2013236","journal-title":"Mol Cell Biol Res Commun"},{"key":"5758_CR40","doi-asserted-by":"publisher","first-page":"1654","DOI":"10.1111\/bph.14171","volume":"175","author":"X Zhu","year":"2018","unstructured":"Zhu X, Finlay DB, Glass M, Duffull SB (2018) An evaluation of the operational model when applied to quantify functional selectivity. Br J Pharmacol 175:1654\u20131668","journal-title":"Br J Pharmacol"},{"key":"5758_CR41","doi-asserted-by":"publisher","first-page":"276","DOI":"10.1016\/j.intimp.2007.10.001","volume":"8","author":"R Zubakova","year":"2008","unstructured":"Zubakova R, Gille A, Faussner A, Hilgenfeldt U (2008) Ca2+ signalling of kinins in cells expressing rat, mouse and human B1\/B2-receptor. Int Immunopharmacol 8:276\u2013281","journal-title":"Int Immunopharmacol"}],"container-title":["Naunyn-Schmiedeberg's Archives of Pharmacology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s00210-026-05758-z.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s00210-026-05758-z","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s00210-026-05758-z.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T01:47:49Z","timestamp":1785808069000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s00210-026-05758-z"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,4]]},"references-count":41,"alternative-id":["5758"],"URL":"https:\/\/doi.org\/10.1007\/s00210-026-05758-z","relation":{},"ISSN":["0028-1298","1432-1912"],"issn-type":[{"value":"0028-1298","type":"print"},{"value":"1432-1912","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,4]]},"assertion":[{"value":"16 April 2026","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"21 July 2026","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"4 August 2026","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","label":"Competing interests","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"Not applicable.","order":2,"name":"Ethics","label":"Clinical trial number","group":{"name":"EthicsHeading","label":"Declarations"}}]}}