A Low Molecular Weight Agonist Signals by Binding to the Transmembrane Domain of Thyroid-stimulating Hormone Receptor (TSHR) and Luteinizing Hormone/Chorionic Gonadotropin Receptor (LHCGR)

Jäschke, H; Neumann, Susanne; Moore, Susanna; Thomas, Craig J.; Colson, Anny‐Odile; Costanzi, Stefano; Kleinau, Gunnar; Jiang, Jian‐kang; Paschke, Ralf; Raaka, Bruce M.; Krause, Gerd; Gershengorn, Marvin C.

doi:10.1074/jbc.c600014200

Cited by 95 publications

(86 citation statements)

References 25 publications

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“…Docking experiments predicted that compound 2 binds to the same cavity in the transmembrane helical bundle of TSHR as previously reported for a smallmolecule partial agonist, Org41841, and an antagonist (14,23), but compound 2 appears to have different sites of interaction (Fig. 2B).…”

Section: Compound 2 Binds To the Transmembrane Helical Bundle Of Tshrmentioning

confidence: 64%

“…We used a TSHR mutant that does not contain the N-terminal ectodomain and that was shown previously not to be activated by TSH (24) and showed that it was activated by compound 2. We also used a 3D homology model of TSHR (14,23), which was based on the x-ray crystallographic structure of opsin (25), to predict the binding pocket for the small-molecule agonist and predict the amino acid residues within the pocket that interact with compound 2. By using a site-specific mutation within the putative TSHR binding pocket, we provided evidence that supports a transmembrane bundle site for the binding of compound 2.…”

Section: Discussionmentioning

confidence: 99%

“…Based on our previous studies of 2 small-molecule ligands of TSHR, a partial agonist Org41841 (23) and an antagonist (14), we predicted that compound 2 would bind to the serpentine region of TSHR. To show that compound 2 binds to the serpentine domain, we tested the activation of a TSHR mutant in which the large amino-terminal ectodomain responsible for TSH binding was deleted.…”

Section: Compound 2 Binds To the Transmembrane Helical Bundle Of Tshrmentioning

confidence: 99%

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Small-molecule agonists for the thyrotropin receptor stimulate thyroid function in human thyrocytes and mice

Neumann¹,

Huang

Titus

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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113

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Seven-transmembrane-spanning receptors (7TMRs) are prominent drug targets. However, small-molecule ligands for 7-transmembrane-spanning receptors for which the natural ligands are large, heterodimeric glycoprotein hormones, like thyroid-stimulating hormone (TSH; thyrotropin), have only recently been reported, and none are approved for human use. We have used quantitative high-throughput screening to identify a small-molecule TSH receptor (TSHR) agonist that was modified to produce a second agonist with increased potency. We show that these agonists are highly selective for human TSHR versus other glycoprotein hormone receptors and interact with the receptor's serpentine domain. A binding pocket within the transmembrane domain was defined by docking into a TSHR homology model and was supported by site-directed mutagenesis. In primary cultures of human thyrocytes, both TSH and the agonists increase mRNA levels for thyroglobulin, thyroperoxidase, sodium iodide symporter, and deiodinase type 2, and deiodinase type 2 enzyme activity. Moreover, oral administration of the agonist stimulated thyroid function in mice, resulting in increased serum thyroxine and thyroidal radioiodide uptake. Thus, we discovered a small molecule that activates human TSHR in vitro, is orally active in mice, and could be a lead for development of drugs to use in place of recombinant human TSH in patients with thyroid cancer.7TMR ͉ G protein-coupled receptor ͉ low-molecular-weight ligands ͉ radioiodide uptake ͉ TSH receptor S mall-molecule agonists and antagonists for 7-transmembrane-spanning receptors (7TMRs; G protein-coupled receptors) make up approximately 40% of the drugs in clinical use (1). The natural ligands for the 7TMRs for which drugs have been discovered are themselves primarily small molecules, such as norepinephrine, adenosine, and acetylcholine. Smallmolecule ligands for the subclass of 7TMRs for which the natural ligands are 30-kDa heterodimeric glycoprotein hormones have recently begun to be described, but none have been approved for use in humans. For example, small-molecule agonists (2-5) and an antagonist (6) for the luteinizing hormone/chorionic gonadotropin receptor (LHCGR), and small-molecule agonists (7-9) and antagonists (10-12) for the FSH receptor (FSHR) have been reported. The thyrotropin (thyroid-stimulating hormone, TSH) receptor (TSHR) is the third member of the glycoprotein hormone receptor subfamily. We recently reported smallmolecule agonists (13) and an antagonist (14) for human TSHR; however, the agonists were of low affinity and were shown to activate TSHRs only in a model cell system in which human TSHRs were ectopically over-expressed.The physiologic role of TSHRs in the hypothalamic-pituitarythyroid axis is well known. TSH, which is produced in the thyrotrophs of the anterior pituitary gland, is secreted into the circulation in response to TSH-releasing hormone stimulation and in turn stimulates the function of the thyroid follicular cells (i.e., thyrocytes) leading, in particular, to increases in size ...

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Section: Compound 2 Binds To the Transmembrane Helical Bundle Of Tshrmentioning

confidence: 64%

Section: Discussionmentioning

confidence: 99%

Section: Compound 2 Binds To the Transmembrane Helical Bundle Of Tshrmentioning

confidence: 99%

See 1 more Smart Citation

Small-molecule agonists for the thyrotropin receptor stimulate thyroid function in human thyrocytes and mice

Neumann¹,

Huang

Titus

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

113

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“…These features are very similar to the group of glycoprotein hormone receptors, such as thyrotropin and lutropin receptors. Mutagenesis studies with these glycoprotein hormone receptors have shown that the large ectodomain is not required for ligand-and mutation-induced signaling of the 7TM domain (41,42).…”

Section: Gpr133 Displays Increased Basal Activity In the G S Protein/mentioning

confidence: 99%

Cell Adhesion Receptor GPR133 Couples to Gs Protein

Bohnekamp

Schöneberg

2011

Journal of Biological Chemistry

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Adhesion G protein-coupled receptors (GPCR), with their very large and complex N termini, are thought to participate in cell-cell and cell-matrix interactions and appear to be highly relevant in several developmental processes. Their intracellular signaling is still poorly understood. Here we demonstrate that GPR133, a member of the adhesion GPCR subfamily, activates the G s protein/adenylyl cyclase pathway. The presence of the N terminus and the cleavage at the GPCR proteolysis site are not required for G protein signaling. G s protein coupling was verified by G␣ s knockdown with siRNA, overexpression of G␣ s , coexpression of the chimeric Gq s4 protein that routes GPR133 activity to the phospholipase C/inositol phosphate pathway, and missense mutation within the transmembrane domain that abolished receptor activity without changing cell surface expression. It is likely that not only GPR133 but also other adhesion GPCR signal via classical receptor/G protein-interaction.Adhesion receptors comprise the second largest subfamily of putatively G protein-coupled receptors (GPCR) 2 with more than 30 members in vertebrates (1, 2). Adhesion GPCR are characterized by long extracellular N termini, which are composed of multiple functional domains, a seven-transmembrane spanning (7TM) domain, and a cytoplasmic tail. Adhesion GPCR are believed to play a role in immune functions (3, 4), angiogenesis (5), cell polarity (6, 7), and development (8, 9). Mutations in some members of the protein family were identified as the cause of inherited developmental defects in humans such as Usher syndrome (VLGR1) (10) and bilateral frontoparietal polymicrogyria (GPR56) (11). Although there is consensus on the fact that this receptor class mediates essential cellcell and cell-matrix interactions (1, 12), the molecular mechanism of intracellular signal transduction of adhesion GPCR remains obscure.There are only a few studies on intracellular signaling mechanisms of adhesion GPCR. Latrophilin 1, the prototype of adhesion GPCR, induces intracellular Ca 2ϩ signaling upon interaction with the exogenous ligand ␣-latrotoxin (13, 14). GPR56 appears to activate the G 12/13 protein/Rho pathway after stimulation with an antibody against the ectodomain (15). BAI1 recognizes phosphatidylserine and can directly recruit a Racguanine nucleotide exchange factor (Rac-GEF) complex to mediate the uptake of apoptotic cells (16). The cytoplasmic domain of BAI2 interacts with GA-binding protein ␥, and GAbinding protein-␣/␥ or GA-binding protein-␣/␤ work as transcriptional repressors of VEGF (17). However, clear evidence of intracellular signaling for most adhesion GPCR via G proteins is still missing (12).Genetic variations in the GPR133 gene, also a member of the adhesion GPCR family, were associated with adult height (18) and the RR interval duration in electrocardiograms (19). GPR133 is expressed in CNS (20) and other tissues; its endogenous agonists and the signal transduction are unknown. Here we demonstrate that GPR133 is coupled to the G s protein/adeny...

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“…This view is borne out in pharmaceutical development by the observation of the relatively more commonly available peptidomimetics for receptors with intramembrane ligand binding, compared with those that bind ligand at the amino terminus. It is true that there are some nonpeptide compounds that bind gonadotropin receptors and produce effects (van Straten et al, 2002), but these are not true competitors at the ligand binding site (which occurs at the large amino-terminal extension); in fact, binding occurs in or near the membrane-associated components of the receptor (Jä schke et al, 2006).…”

Section: B Lessons From Comparison Of the Gonadotropinreleasing Hormmentioning

confidence: 99%

G Protein-Coupled Receptor Trafficking in Health and Disease: Lessons Learned to Prepare for Therapeutic Mutant Rescue in Vivo

2007

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A Low Molecular Weight Agonist Signals by Binding to the Transmembrane Domain of Thyroid-stimulating Hormone Receptor (TSHR) and Luteinizing Hormone/Chorionic Gonadotropin Receptor (LHCGR)

Cited by 95 publications

References 25 publications

Small-molecule agonists for the thyrotropin receptor stimulate thyroid function in human thyrocytes and mice

Small-molecule agonists for the thyrotropin receptor stimulate thyroid function in human thyrocytes and mice

Cell Adhesion Receptor GPR133 Couples to Gs Protein

G Protein-Coupled Receptor Trafficking in Health and Disease: Lessons Learned to Prepare for Therapeutic Mutant Rescue in Vivo

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