Parallel bioassay of 39 tachykinins on 11 smooth muscle preparations. Structure and receptor selectivity/affinity relationship

Severini, Cinzia; Salvadori, Severo; Guerrini, Remo; Falconieri–Erspamer, Giuliana; Mignogna, Giuseppina; Erspamer, V.

doi:10.1016/s0196-9781(00)00290-4

Cited by 47 publications

(53 citation statements)

References 31 publications

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“…The NK1 receptor is one of three tachykinin receptor subtypes, whose endogenous ligand is the Substance P peptide. The NK1 receptor is distributed in the central and peripheral nervous system and is implicated in depression, asthma, emesis, anxiety, and pain (43). There is only one marketed drug that targets this receptor, aprepitant (Emend), which is approved for emesis.…”

Section: Resultsmentioning

confidence: 99%

G protein-coupled receptors:In silicodrug discovery in 3D

Becker¹,

Marantz²,

Shacham³

et al. 2004

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

164

158

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The application of structure-based in silico methods to drug discovery is still considered a major challenge, especially when the x-ray structure of the target protein is unknown. Such is the case with human G protein-coupled receptors (GPCRs), one of the most important families of drug targets, where in the absence of x-ray structures, one has to rely on in silico 3D models. We report repeated success in using ab initio in silico GPCR models, generated by the PREDICT method, for blind in silico screening when applied to a set of five different GPCR drug targets. More than 100,000 compounds were typically screened in silico for each target, leading to a selection of <100 ''virtual hit'' compounds to be tested in the lab. In vitro binding assays of the selected compounds confirm high hit rates, of 12-21% (full dose-response curves, K i < 5 M). In most cases, the best hit was a novel compound (New Chemical Entity) in the 1-to 100-nM range, with very promising pharmacological properties, as measured by a variety of in vitro and in vivo assays. These assays validated the quality of the hits as lead compounds for drug discovery. The results demonstrate the usefulness and robustness of ab initio in silico 3D models and of in silico screening for GPCR drug discovery.modeling ͉ in silico screening ͉ structure-based G protein-coupled receptors (GPCRs) are membraneembedded proteins, responsible for communication between the cell and its environment (1). As a consequence, many major diseases, such as hypertension, cardiac dysfunction, depression, anxiety, obesity, inflammation, and pain, involve malfunction of these receptors (2), making them among the most important drug targets for pharmacological intervention (3-5). Thus, whereas GPCRs are only a small subset of the human genome, they are the targets for Ϸ50% of all recently launched drugs (6). As targets of paramount importance, it is expected that drug discovery for GPCRs would benefit from the introduction of computational methodologies (7), especially as these methods can be used in conjunction with such experimental methods as high-throughput screening (8, 9), NMR, and crystallography (10).Unfortunately, GPCRs, like other membrane-embedded proteins, have characteristics that make their 3D structure extremely difficult to determine experimentally. To date, the only GPCR for which a 3D structure was determined by x-ray crystallography is bovine rhodopsin (11), which is unique among GPCRs in that its ligand, retinal, is covalently bound and that it responds to light rather than to ligand binding. Hence, in the case of GPCRs, the limited availability of structural data has forced the computational design of ligands to heavily rely on ligand-based techniques. Indeed, for many GPCRs, the natural ligand can provide a good starting point, leading to useful pharmacophore models that can be used for identifying lead structures with novel scaffolds (6). These methods have been successfully applied for the discovery of peptide agonists to the somatostatin receptor (12) and for...

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Section: Resultsmentioning

confidence: 99%

G protein-coupled receptors:In silicodrug discovery in 3D

Becker¹,

Marantz²,

Shacham³

et al. 2004

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

164

158

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“…7D). Intriguingly, YSEC, but not MAE cells, expressed NK3, which binds NK-B with higher affinity than other NK receptors (47). Thus, YSEC express functional NK receptors, which can mediate activation of adenylyl cylase in response to NK-B.…”

Section: Gata-1-mediated Activation Of Neurokinin-b Transcriptionmentioning

confidence: 94%

“…In other scenarios, Nab1 and Nab2 are co-regulated. 3 Tac-2 is highly related to Tac-1, which encodes preprotachykinin-1, the precursor to Substance P that mediates pain perception (47). To assess the specificity of GATA-1-mediated repression of Nab2 and activation of Tac-2, we tested whether ER-GATA-1 regulates Nab1 and Tac-1 expression.…”

Section: Gata-1-mediated Activation Of Neurokinin-b Transcriptionmentioning

confidence: 99%

“…An Erythroid Cell-derived Neurokinin: Implications for Vascular Regulation-Whereas neurokinins were classically studied in the nervous system, recent studies have expanded the repertoire of sites of neurokinin synthesis and function (47,50). However, whether erythroid cells are competent to generate neurokinins had not been investigated.…”

Section: Gata-1-mediated Activation Of Neurokinin-b Transcriptionmentioning

confidence: 99%

“…To assess whether NK-B functions via autocrine or paracrine regulation of erythroid progenitors, we tested whether the G protein-coupled NK receptors (NK1, NK2, and NK3) that mediate NK-B actions (57) are expressed in uninduced or induced G1E-ER-GATA-1 cells. NK-B has the highest affinity for NK3 but can also bind and activate NK1 and NK2 (47). NK receptor activation results in stimulation of adenylyl cyclase and cyclic AMP generation, as well as activation of phospholipase C and the accumulation of intracellular calcium (58,59).…”

Section: Gata-1-mediated Activation Of Neurokinin-b Transcriptionmentioning

confidence: 99%

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Neurokinin-B Transcription in Erythroid Cells

Pal

Nemeth

Bodine

et al. 2004

Journal of Biological Chemistry

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The GATA family of transcription factors establishes genetic networks that control developmental processes including hematopoiesis, vasculogenesis, and cardiogenesis. We found that GATA-1 strongly activates transcription of the Tac-2 gene, which encodes proneurokinin-B, a precursor of neurokinin-B (NK-B). Neurokinins function through G protein-coupled transmembrane receptors to mediate diverse physiological responses including pain perception and the control of vascular tone. Whereas an elevated level of NK-B was implicated in pregnancy-associated pre-eclampsia (Page, N. M., Woods, R. J., Gardiner, S. M., Lomthaisong, K., Gladwell, R. T., Butlin, D. J., Manyonda, I. T., and Lowry, P. J. (2000) Nature 405, 797-800), the regulation of NK-B synthesis and function are poorly understood. Tac-2 was expressed in normal murine erythroid cells and was induced upon ex vivo erythropoiesis. An estrogen receptor fusion to GATA-1 (ER-GATA-1) and endogenous GATA-1 both occupied a region of Tac-2 intron-7, which contains two conserved GATA motifs. Genetic complementation analysis in GATA-1-null G1E cells revealed that endogenous GATA-2 occupied the same region of intron-7, and expression of ER-GATA-1 displaced GATA-2 and activated Tac-2 transcription. Erythroid cells did not express neurokinin receptors, whereas aortic and yolk sac endothelial cells differentially expressed neurokinin receptor subtypes. Since NK-B induced cAMP accumulation in yolk sac endothelial cells, these results suggest a new mode of vascular regulation in which GATA-1 controls NK-B synthesis in erythroid cells.The development of blood cells from hematopoietic stem cells is critically dependent upon integration of the activities of diverse transcription factors. Of particular importance are three members of the GATA family of transcription factors that regulate hematopoiesis via both unique and overlapping activities (1). GATA-1 is required for definitive or adult erythropoiesis, for megakaryocyte maturation, and for eosinophil production (2-7), whereas GATA-2 is important for the survival and function of multipotent hematopoietic precursor cells (8 -10). GATA-1 and GATA-2 have redundant activities to control primitive or embryonic erythropoiesis (11). GATA-3 uniquely regulates aspects of lymphopoiesis and central nervous system development (12-16). Understanding how GATA factors regulate development requires the elucidation of genetic circuits that control expression of GATA factor target genes and the GATA factors themselves.The C-terminal zinc finger of dual zinc finger GATA factors recognizes a simple DNA motif (WGATAR) (17-20) that is distributed abundantly throughout genomes. In contrast, the N-terminal zinc finger mediates interactions with the friend of GATA-1 (FOG-1) 1 coregulator (21, 22) and stabilizes DNA binding at certain GATA motifs (17,23). Despite the high frequency of WGATAR in genomes, analyses of GATA-1 and GATA-2 binding to chromatin by quantitative ChIP analysis have revealed an exquisite specificity of chromatin occupancy. For exampl...

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Solution conformation of non-mammalian tachykinin physalaemin in lipid micelles by nuclear magnetic resonance

Grace

Cowsik

2011

Biopolymers

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Physalaemin (PHY), a non-mammalian tachykinin, binds selectively to neurokinin-1 (NK1) receptor with high affinity. Both the aqueous and lipid-induced conformations of PHY have been studied using two-dimensional nuclear magnetic resonance techniques. These data show that in water PHY prefers to be in an extended conformation and that in the presence of perdeuterated dodecylphosphocholine micelles, a membrane model system, a helical conformation is observed from Pro4 to the C-terminus. Comparison of the structures ofPHYand other NK ligands along with structure activity studies reported on these peptide ligands suggests that helical backbone structural motif is necessary for the binding of these NK ligands to the various NK receptors. Furthermore, consensus in the structures of these ligands suggests that these ligands must be binding along the highly hydrophobic face of the helix that contains the important hydrophobic residues, Phe7, Leu10, and Met11, that are highly conserved in most of the ligands.

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Parallel bioassay of 39 tachykinins on 11 smooth muscle preparations. Structure and receptor selectivity/affinity relationship

Cited by 47 publications

References 31 publications

G protein-coupled receptors:In silicodrug discovery in 3D

G protein-coupled receptors:In silicodrug discovery in 3D

Neurokinin-B Transcription in Erythroid Cells

Solution conformation of non-mammalian tachykinin physalaemin in lipid micelles by nuclear magnetic resonance

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