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Cited by 65Receptor Mechanisms and SignalingThyroid Disorders and TreatmentsProtein Kinase Regulation and GTPase Signaling

Occupancy of both sites on the thyrotropin (TSH) receptor dimer is necessary for phosphoinositide signaling

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Allen, Michael D., Neumann, Susanne, Gershengorn, Marvin C.·2011·The FASEB Journal
DOI 10.1096/fj.11-188961PMC PMC3177577
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Abstract

The thyroid‐stimulating hormone (TSH) receptor signals via G s to produce cAMP and via G q/11 to produce inositol‐1,4,5‐trisphosphate, which is degraded to inositol monophosphate (IP1; phosphoinositide signaling). The potency of TSH for cAMP signaling is higher than for phosphoinositide signaling, and it was suggested that there are “spare receptors” for cAMP signaling. In a human embryonic kidney macrophage scavenger receptor‐expressing (HEK‐EM) 293 model system, there are no spare receptors, but the cells still exhibited 100‐fold differences in potencies. Dose responses for TSH‐stimulated dissociation of prebound 125 I‐TSH (negative cooperativity; EC 50 =70 mU/ml), which requires TSH binding to both sites of the TSH receptor (TSHR) homodimer, and TSH‐stimulated IP1 production (EC 50 =50 mU/ml) were indistinguishable. Fluorescence resonance energy transfer (FRET) using tagged receptors showed that TSHR formed homodimers and heterodimers with two binding‐deficient mutant TSHRs, L252P and C41S. When L252P or C41S was expressed with TSHR, that is, when TSHR/L252P or TSHR/C41S heterodimers could only bind one TSH, TSH‐stimulated IP1 production was decreased relative to cAMP production. The slopes of linear regression analyses comparing fold stimulation by TSH of IP1 vs. cAMP production were 0.044 ± 0.0047, 0.0043 ± 0.0041, and 0.0059 ± 0.0014 for cells expressing TSHR alone, TSHR and L252P, or TSHR and C41S, respectively. We suggest that TSHR coupling to phosphoinositide signaling is dependent on binding 2 molecules of TSH to TSHR homodimer, causing a conformational change allowing coupling to G q/11 .—Allen, M. D., Neumann, S., Gershengorn, M. C. Occupancy of both sites on the thyrotropin (TSH) receptor dimer is necessary for phosphoinositide signaling. FASEB J. 25, 3687–3694 (2011). www.fasebj.org

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  • Low-concentration TSH binds to the high-affinity binding site on one of the protomers of the TSHR homodimer, and preferentially activates Gαs to stimulate cAMP accumulation, which then activates PKA.

    “Low-concentration TSH binds to the high-affinity binding site on one of the protomers of the TSHR homodimer, and preferentially activates Gαs to stimulate cyclic adenosine monophosphate (cAMP) accumulation, which then activates protein kinase A (PKA) (120).”

    From Intrathyroidal feedforward and feedback network regulating thyroid hormone synthesis and secretion (2022) · cites this paper as [120]

    The Discussion (page not available) says TSH binding at the high-affinity site on one TSHR homodimer protomer favors Gs activation and stimulates cAMP production; the Results also show cAMP signaling at lower TSH concentrations than phosphoinositide signaling. Our copy does not state that the resulting cAMP activates PKA, so that final step is not supported by this source. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • High-concentration TSH binds to both the high- and low-affinity protomers, activating both Gαs and Gαq/11 to increase cAMP production and activate PKC, respectively.

    “High-concentration TSH binds to both the high- and low-affinity protomers, activating both Gαs and Gαq/11 to increase cAMP production and activate PKC, respectively (120).”

    From Intrathyroidal feedforward and feedback network regulating thyroid hormone synthesis and secretion (2022) · cites this paper as [120]

    The Discussion states that TSH occupancy of both the high- and low-affinity sites permits Gs and Gq/11 activation, increasing cAMP and IP1 production. The paper describes phosphoinositide signaling through Gq/11 but does not report PKC activation; that part of the attributed claim is unsupported. (LLM, conf=0.89)

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  1. [1]1. Laugwitz K. L., Allgeier A., Offermanns S., Spicher K., Van Sande J., Dumont J. E., Schultz G. (1996) The human thyrotropin receptor: A heptahelical receptor capable of stimulating members of all four G protein families. Proc. Natl. Acad. Sci. U. S. A. 93, 116–120 [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. [2]2. Corda D., Marcocci C., Kohn L. D., Axelrod J., Luini A. (1985) Association of the changes in cytosolic Ca2+ and iodide efflux induced by thyrotropin and by the stimulation of alpha 1-adrenergic receptors in cultured rat thyroid cells. J. Biol. Chem. 260, 9230–9236 [PubMed] [Google Scholar]
  3. [3]3. Nussenzveig D. R., Thaw C. N., Gershengorn M. C. (1994) Inhibition of inositol phosphate second messenger formation by intracellular loop one of a human calcitonin receptor. Expression and mutational analysis of synthetic receptor genes. J. Biol. Chem. 269, 28123–28129 [PubMed] [Google Scholar]
  4. [4]4. Offermanns S., Iida-Klein A., Segre G. V., Simon M. I. (1996) Gαq family members couple parathyroid hormone (PTH)/PTH-related peptide and calcitonin receptors to phospholipase C in COS-7 cells. Mol. Endocrinol. 10, 566–574 [DOI] [PubMed] [Google Scholar]
  5. [5]5. Trimble E. R., Bruzzone R., Biden T. J., Farese R. V. (1986) Secretin induces rapid increases in inositol trisphosphate, cytosolic Ca2+ and diacylglycerol as well as cyclic AMP in rat pancreatic acini. Biochem. J. 239, 257–261 [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. [6]6. Kuhn B., Schmid A., Harteneck C., Gudermann T., Schultz G. (1996) G proteins of the Gq family couple the H2 histamine receptor to phospholipase C. Mol. Endocrinol. 10, 1697–1707 [DOI] [PubMed] [Google Scholar]
  7. [7]7. Latif R., Graves P., Davies T. F. (2001) Oligomerization of the human thyrotropin receptor. Fluorescent protein-tagged hTSHR reveals post-translational complexes. J. Biol. Chem. 276, 45217–45224 [DOI] [PubMed] [Google Scholar]
  8. [8]8. Gao F., Harikumar K. G., Dong M., Lam P. C., Sexton P. M., Christopoulos A., Bordner A., Abagyan R., Miller L. J. (2009) Functional importance of a structurally distinct homodimeric complex of the family B G protein-coupled secretin receptor. Mol. Pharmacol. 76, 264–274 [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. [9]9. De Meyts P., Gauguin L., Svendsen A. M., Sarhan M., Knudsen L., Nohr J., Kiselyov V. V. (2009) Structural basis of allosteric ligand-receptor interactions in the insulin/relaxin peptide family: implications for other receptor tyrosine kinases and G-protein-coupled receptors. Ann. N. Y. Acad. Sci. 1160, 45–53 [DOI] [PubMed] [Google Scholar]
  10. [10]10. Urizar E., Montanelli L., Loy T., Bonomi M., Swillens S., Gales C., Bouvier M., Smits G., Vassart G., Costagliola S. (2005) Glycoprotein hormone receptors: link between receptor homodimerization and negative cooperativity. EMBO J. 24, 1954–1964 [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. [11]11. Gurevich V. V., Gurevich E. V. (2008) How and why do GPCRs dimerize? Trends Pharmacol. Sci. 29, 234–240 [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. [12]12. Han Y., Moreira I. S., Urizar E., Weinstein H., Javitch J. A. (2009) Allosteric communication between protomers of dopamine class A GPCR dimers modulates activation. Nat. Chem. Biol. 5, 688–695 [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. [13]13. Milligan G. (2009) G protein-coupled receptor hetero-dimerization: contribution to pharmacology and function. Br. J. Pharmacol. 158, 5–14 [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. [14]14. Pin J. P., Neubig R., Bouvier M., Devi L., Filizola M., Javitch J. A., Lohse M. J., Milligan G., Palczewski K., Parmentier M., Spedding M. (2007) International Union of Basic and Clinical Pharmacology. LXVII. Recommendations for the recognition and nomenclature of G protein-coupled receptor heteromultimers. Pharmacol. Rev. 59, 5–13 [DOI] [PubMed] [Google Scholar]
  15. [15]15. Lee S. P., So C. H., Rashid A. J., Varghese G., Cheng R., Lanca A. J., O'Dowd B. F., George S. R. (2004) Dopamine D1 and D2 receptor Co-activation generates a novel phospholipase C-mediated calcium signal. J. Biol. Chem. 279, 35671–35678 [DOI] [PubMed] [Google Scholar]
  16. [16]16. Shaner N. C., Campbell R. E., Steinbach P. A., Giepmans B. N., Palmer A. E., Tsien R. Y. (2004) Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nat. Biotechnol. 22, 1567–1572 [DOI] [PubMed] [Google Scholar]
  17. [17]17. Tonacchera M., Perri A., De Marco G., Agretti P., Banco M. E., Di Cosmo C., Grasso L., Vitti P., Chiovato L., Pinchera A. (2004) Low prevalence of thyrotropin receptor mutations in a large series of subjects with sporadic and familial nonautoimmune subclinical hypothyroidism. J. Clin. Endocrinol. Metab. 89, 5787–5793 [DOI] [PubMed] [Google Scholar]
  18. [18]18. De Roux N., Misrahi M., Brauner R., Houang M., Carel J. C., Granier M., Le Bouc Y., Ghinea N., Boumedienne A., Toublanc J. E., Milgrom E. (1996) Four families with loss of function mutations of the thyrotropin receptor. J. Clin. Endocrinol. Metab. 81, 4229–4235 [DOI] [PubMed] [Google Scholar]
  19. [19]19. Neumann S., Huang W., Titus S., Krause G., Kleinau G., Alberobello A. T., Zheng. W., Southall N., Inglese J., Austin C. P., Celi F. S., Gavrilova O., Thomas C. J., Raaka B. M., Gershengorn M. C. (2009) Small-molecule agonists for the thyrotropin receptor stimulate thyroid function in human thyrocytes and mice. Proc. Natl. Acad. Sci. U. S. A. 106, 12471–12476 [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. [20]20. Deflorian F., Engel S., Colson A. O., Raaka B. M., Gershengorn M. C., Costanzi S. (2008) Understanding the structural and functional differences between mouse thyrotropin-releasing hormone receptors 1 and 2. Proteins 71, 783–794 [DOI] [PubMed] [Google Scholar]
  21. [21]21. Dalrymple M. B., Pfleger K. D., Eidne K. A. (2008) G protein-coupled receptor dimers: functional consequences, disease states and drug targets. Pharmacol. Ther. 118, 359–371 [DOI] [PubMed] [Google Scholar]
  22. [22]22. Rivero-Muller A., Chou Y. Y., Ji I., Lajic S., Hanyaloglu A. C., Jonas K., Rahman N., Ji T. H., Huhtaniemi I. (2010) Rescue of defective G protein-coupled receptor function in vivo by intermolecular cooperation. Proc. Natl. Acad. Sci. U. S. A. 107, 2319–2324 [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. [23]23. Ernst O. P., Gramse V., Kolbe M., Hofmann K. P., Heck M. (2007) Monomeric G protein-coupled receptor rhodopsin in solution activates its G protein transducin at the diffusion limit. Proc. Natl. Acad. Sci. U. S. A. 104, 10859–10864 [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. [24]24. Whorton M. R., Jastrzebska B., Park P. S., Fotiadis D., Engel A., Palczewski K., Sunahara R. K. (2008) Efficient coupling of transducin to monomeric rhodopsin in a phospholipid bilayer. J. Biol. Chem. 283, 4387–4394 [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. [25]25. Springael J. Y., Urizar E., Costagliola S., Vassart G., Parmentier M. (2007) Allosteric properties of G protein-coupled receptor oligomers. Pharmacol. Ther. 115, 410–418 [DOI] [PubMed] [Google Scholar]
  26. [26]26. White J. F., Grodnitzky J., Louis J. M., Trinh L. B., Shiloach J., Gutierrez J., Northup J. K., Grisshammer R. (2007) Dimerization of the class A G protein-coupled neurotensin receptor NTS1 alters G protein interaction. Proc. Natl. Acad. Sci. U. S. A. 104, 12199–12204 [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. [27]27. Damian M., Martin A., Mesnier D., Pin J. P., Baneres J. L. (2006) Asymmetric conformational changes in a GPCR dimer controlled by G-proteins. EMBO J. 25, 5693–5702 [DOI] [PMC free article] [PubMed] [Google Scholar] Associated Data This section collects any data citations, data availability statements, or supplementary materials included in this article.

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Authors
Allen, M. D., Neumann, S., & Gershengorn, M. C.
Affiliations
National Institutes of Health (US)
Corresponding author
Gershengorn, Marvin C.
ORCID
Venue
The FASEB Journal
Publisher
Wiley
Published
2011-06-24
Volume
25
Issue
10
Pages
3687-3694
ISSN
  • 0892-6638 (ISSN-L)
  • 1530-6860
DOI
10.1096/fj.11-188961
PMC
PMC3177577
OpenAlex
W2156167242
Cited by
65
References
0
Citations by year
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FWCI
1.69
Type
journal article
Language
en
Open access
Licence
onlinelibrary.wiley.com/termsAndConditions#vor (vor)
Locations
  • The FASEB Journal (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
National Institutes of Health
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
  • Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Thyroid Disorders and Treatments
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
Keywords
TSH receptor, phosphoinositide signaling, inositol 1,4,5-trisphosphate, inositol monophosphate, cAMP signaling, Gq/11 coupling, Gs coupling, receptor heterodimers, negative cooperativity, receptor conformational change, spare receptors
MeSH terms
Cyclic AMP, Gene Expression Regulation, Humans, Phosphatidylinositols, Protein Binding, Receptors, Thyrotropin, Thyrotropin, Signal Transduction, GTP-Binding Protein alpha Subunits, Gs, Protein Subunits, GTP-Binding Protein alpha Subunits, Gq-G11, HEK293 Cells

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