Stereospecific, high affinity binding of 2,3,7,8-tetrachlorodibenzo-p-dioxin by hepatic cytosol. Evidence that the binding species is receptor for induction of aryl hydrocarbon hydroxylase.

Poland, Alan; Glover, Edward; Kende, Andrew S.

doi:10.1016/s0021-9258(17)33205-2

Cited by 1,243 publications

(131 citation statements)

References 24 publications

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“…Though the obtained binding affinities for the individually tested ligands to the recombinant protein are slightly lower than the ones reported in the literature for the hAhR [12,25,27], such differences are presumably explainable by missing endogenous factors that are likely to support ligand sensing of the cytosolic AhR, e.g., stabilizing effects of chaperones (HSP90, XAP2) in the unbound receptor state [30,31]. Moreover, and of note, most of the literature-described binding affinities were experimentally determined for the murine AhR, which is known to display an almost 10-fold higher binding affinity towards TCDD than human AhR [32].…”

Section: Discussionmentioning

confidence: 68%

“…However, not only because of the undesirable usage of radioactive and toxic tracer material, the use of animals for organ extraction and the increased costs involved but also due to diverse technical reasons, there is a necessity for the development of an improved ligand binding assay. Besides the apparent sensitivity towards amendments in the protocol, some studies stress the fact that the radioligand binding assay largely suffers from the insolubility of TCDD and other AhR ligands in water [26,27]. Due to their lipophilic nature, these compounds are usually dissolved in DMSO.…”

Section: Discussionmentioning

confidence: 99%

“…The radioligand binding assay, although commonly used, is known to be extremely sensitive to minor changes in the protocol that likely lead to marked differences in the measured affinities [12,20,27]. Here, total cytosolic extracts of hepatocytes are being used; therefore, the observed ligand binding may not be specific to AhR, but potentially also arise from unspecific interactions of the ligand with other constituents of the cytosol.…”

Section: Mst Is a Sensitive Methods To Determine Ahr-ligand Binding Afmentioning

confidence: 99%

“…However, in agreement with the data obtained from our radioligand saturation experiments, we observed only an initial competition of unlabeled TCDD for binding to hAhR at the highest concentration. Full displacement of the radioligand would require the use of even higher concentrations of unlabeled TCDD, which due to its insolubility in water [27] is not feasible in this experimental setup.…”

Section: Recombinant Ahr-arnt Is Functionally Activementioning

confidence: 99%

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Novel Method for Quantifying AhR-Ligand Binding Affinities Using Microscale Thermophoresis

et al. 2021

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The aryl hydrocarbon receptor (AhR) is a highly conserved cellular sensor of a variety of environmental pollutants and dietary-, cell- and microbiota-derived metabolites with important roles in fundamental biological processes. Deregulation of the AhR pathway is implicated in several diseases, including autoimmune diseases and cancer, rendering AhR a promising target for drug development and host-directed therapy. The pharmacological intervention of AhR processes requires detailed information about the ligand binding properties to allow specific targeting of a particular signaling process without affecting the remaining. Here, we present a novel microscale thermophoresis-based approach to monitoring the binding of purified recombinant human AhR to its natural ligands in a cell-free system. This approach facilitates a precise identification and characterization of unknown AhR ligands and represents a screening strategy for the discovery of potential selective AhR modulators.

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Section: Discussionmentioning

confidence: 68%

Section: Discussionmentioning

confidence: 99%

Section: Mst Is a Sensitive Methods To Determine Ahr-ligand Binding Afmentioning

confidence: 99%

Section: Recombinant Ahr-arnt Is Functionally Activementioning

confidence: 99%

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Novel Method for Quantifying AhR-Ligand Binding Affinities Using Microscale Thermophoresis

et al. 2021

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“…The AhR is a ligand-activated transcription factor and was first described as a receptor for 2,3,7,8-tetrachlorodibenzo-pdioxin (TCDD) (1). Additional exogenous chemicals like benzo (a)pyrene (BaP) but also endogenous ligands like the lightdependent Tryptophan metabolite 6-formylindolo [3,2-b] carbazole (FICZ) and metabolites produced by the intestinal microbiota have been shown to activate the AhR (12,13).…”

Section: Introductionmentioning

confidence: 99%

Non-Genomic AhR-Signaling Modulates the Immune Response in Endotoxin-Activated Macrophages After Activation by the Environmental Stressor BaP

et al. 2021

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Emerging studies revealed that the Aryl hydrocarbon receptor (AhR), a receptor sensing environmental contaminants, is executing an immunomodulatory function. However, it is an open question to which extent this is achieved by its role as a transcription factor or via non-genomic signaling. We utilized a multi-post-translational modification-omics approach to examine non-genomic AhR-signaling after activation with endogenous (FICZ) or exogenous (BaP) ligand in endotoxin-activated (LPS) monocyte-derived macrophages. While AhR activation affected abundances of few proteins, regulation of ubiquitination and phosphorylation were highly pronounced. Although the number and strength of effects depended on the applied AhR-ligand, both ligands increased ubiquitination of Rac1, which participates in PI3K/AKT-pathway-dependent macrophage activation, resulting in a pro-inflammatory phenotype. In contrast, co-treatment with ligand and LPS revealed a decreased AKT activity mediating an anti-inflammatory effect. Thus, our data show an immunomodulatory effect of AhR activation through a Rac1ubiquitination-dependent mechanism that attenuated AKT-signaling, resulting in a mitigated inflammatory response.

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Dibenzo‐ p ‐Dioxins: 2,3,7,8‐Tetrachlorodibenzo‐ p ‐Dioxin and Other Polychlorinated Dioxin‐Like Chemicals

Birnbaum,

De Vito

2023

Patty's Toxicology

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2,3,7,8‐Tetrachlordibenzo‐ p ‐dioxin (TCDD) is often called the most potent toxic man‐made chemical. TCDD is the most toxic member of the polychlorinated dibenzo‐ p ‐dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and coplanar PCBs or “dioxin‐like” chemicals (DLCs). These chemicals have similar structures and toxicological effects. TCDD is considered the prototype of the DLCs due to its toxic potency, persistence in the environment and biological systems and because it is the most studied of the DLCs. While polychlorinated dibenzo‐ p ‐dioxins are a class of compounds with 75 possible congeners, as shown in the following structure, only those chlorinated in all the lateral positions are considered dioxins or DLCs. The lateral chlorines result in planar compounds and impart metabolic stability, biological potency, and environmental persistence. TCDD and other DLCs induce a broad range of biochemical and toxic responses. These effects are elicited through their binding and activating the Ah‐receptor (AhR), which is an intracellular protein that acts as a ligand‐activated transcription factor. The binding and activation of the AhR by DLCs initiate a cascade of biochemical, cellular, tissue, and organ‐level effects leading to developmental toxicity, immunotoxicity, reproductive toxicity, and carcinogenicity, among others.

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Stereospecific, high affinity binding of 2,3,7,8-tetrachlorodibenzo-p-dioxin by hepatic cytosol. Evidence that the binding species is receptor for induction of aryl hydrocarbon hydroxylase.

Cited by 1,243 publications

References 24 publications

Novel Method for Quantifying AhR-Ligand Binding Affinities Using Microscale Thermophoresis

Novel Method for Quantifying AhR-Ligand Binding Affinities Using Microscale Thermophoresis

Non-Genomic AhR-Signaling Modulates the Immune Response in Endotoxin-Activated Macrophages After Activation by the Environmental Stressor BaP

Dibenzo‐ p ‐Dioxins: 2,3,7,8‐Tetrachlorodibenzo‐ p ‐Dioxin and Other Polychlorinated Dioxin‐Like Chemicals

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