Bacterial biosensors for evaluating potential impacts of estrogenic endocrine disrupting compounds in multiple species

Gierach, Izabela; Shapero, Kayle; Eyster, Thomas W.; Wood, David

doi:10.1002/tox.20708

Cited by 15 publications

(23 citation statements)

References 58 publications

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“…Initial studies used the human estrogen (ER␣) and thyroid hormone (TR␤-1) receptors and showed that this sensor can differentiate between receptor agonists and antagonists (71). Further, the modular design of the sensor has simplified the development of additional biosensors based on estrogen (ER␣ and ER␤) (69) and thyroid hormone (TR␣-1 and TR␤-1) receptors (72), as well as the human peroxisome proliferator-activated receptor ␥ (PPAR␥) receptor (73) and several animal estrogen receptors (74).…”

Section: Detecting Small Moleculesmentioning

confidence: 99%

Intein Applications: From Protein Purification and Labeling to Metabolic Control Methods

Wood

Camarero

2014

Journal of Biological Chemistry

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124

102

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The discovery of inteins in the early 1990s opened the door to a wide variety of new technologies. Early engineered inteins from various sources allowed the development of self-cleaving affinity tags and new methods for joining protein segments through expressed protein ligation. Some applications were developed around native and engineered split inteins, which allow protein segments expressed separately to be spliced together in vitro. More recently, these early applications have been expanded and optimized through the discovery of highly efficient trans-splicing and trans-cleaving inteins. These new inteins have enabled a wide variety of applications in metabolic engineering, protein labeling, biomaterials construction, protein cyclization, and protein purification.The ability of inteins to form and cleave specific peptide bonds in a variety of contexts has enabled the development of powerful new tools in molecular biology. Initial applications focused on self-cleaving affinity tags and protein modification and labeling, and utilized mutations that altered the native splicing reactions described by Perler and co-workers (89) in this series. An important advantage of intein-based methods is that they are generally enzymatic in nature, and therefore exhibit highly specific activities under physiological conditions. Although thiol compounds are used in some applications, the chemistries involved in most intein methods are innocuous to their various target proteins. Intein-based methods have greatly expanded in subsequent years through the discovery of hundreds of additional inteins, and new applications in protein activity regulation and modification have followed. An important underlying theme in this work has been the discovery and development of highly efficient split inteins for advanced applications in vitro and in vivo. These trans-splicing and -cleaving inteins have been employed for applications ranging from the purification of recombinant products to the in vivo control of protein function and labeling. Protein PurificationOne of the first major applications of inteins was the development of self-cleaving affinity tags for the recovery of untagged target proteins in recombinant expression systems (1-3). In these applications, a modified intein is expressed in fusion to an affinity tag and target protein, and once the fusion is affinity-purified, the intein is induced to cleave the target protein from the intein and tag (Fig. 1). The first commercial intein system was released by New England Biolabs in 1997 and employed a modified Sce VMA1 3 intein that is triggered to cleave at its N terminus (IMPACT system), or both N and C termini (IMPACT-CN system), by the addition of thiol compounds (e.g. Refs. 4 -7) (Fig. 1A, left). Shortly thereafter, inteins with rapid C-terminal cleaving activity were published. These inteins exhibit suppressed cleavage activity at pH 8.5, allowing purification of the tagged target, and are then induced to cleave by a shift to pH 6.5. The cleavage activity of these inteins is als...

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Section: Detecting Small Moleculesmentioning

confidence: 99%

Intein Applications: From Protein Purification and Labeling to Metabolic Control Methods

Wood

Camarero

2014

Journal of Biological Chemistry

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124

102

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“…It could differentiate agonist from antagonist and give a rough indication of binding affi nity via dose-response curves. In tests the strains correctly identifi ed estrogenic test compounds, so it can be useful in initial comparative analysis of EDCs impacts across multiple species [16].…”

Section: Bacterial and Yeast Biosensors For Evaluating Potential Impamentioning

confidence: 99%

“…The biological activity of EDCs have been monitored by use of biomarkers such as vitellogenin, choliogenin and a DNA binding assay. Because of their diversity and simple nature, genetically engineered bioluminescent E. coli biosensors, which are sensitive to oxidative, DNA, membrane or protein damage, have been applied to EDCs screening and classifi cation [8,16]. Applications of reporter genes for screening of hormonal active compounds in the environment are broadly described and summarized by Svobodová and Cajthaml 2010 [41].…”

Section: Bacterial and Yeast Biosensors For Evaluating Potential Impamentioning

confidence: 99%

“…al., 2011 [16] developed bacterial biosensors for evaluating potential impacts of estrogenic endocrine disrupting compounds in multiple species. Escherichia coli were incorporated with the ligand-binding domains (LBDs) of the β-subtype estrogen receptors (ERβ) from Solea solea (sole), and Sus scrofa (pig).…”

Section: Bacterial and Yeast Biosensors For Evaluating Potential Impamentioning

confidence: 99%

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Environmental Application of Reporter-Genes Based Biosensors for Chemical Contamination Screening

Matejczyk¹,

Rosochacki²

2014

Archives of Environmental Protection

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Abstract:The paper presents results of research concerning possibilities of applications of reporter-genes based microorganisms, including the selective presentation of defects and advantages of different new scientifi c achievements of methodical solutions in genetic system constructions of biosensing elements for environmental research. The most robust and popular genetic fusion and new trends in reporter genes technology -such as LacZ (β-galactosidase), xylE (catechol 2,3-dioxygenase), gfp (green fl uorescent proteins) and its mutated forms, lux (prokaryotic luciferase), luc (eukaryotic luciferase), phoA (alkaline phosphatase), gusA and gurA (β-glucuronidase), antibiotics and heavy metals resistance are described. Reporter-genes based biosensors with use of genetically modifi ed bacteria and yeast successfully work for genotoxicity, bioavailability and oxidative stress assessment for detection and monitoring of toxic compounds in drinking water and different environmental samples, surface water, soil, sediments.

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“…In particular, the binding of an appropriate ligand to the fused LBD activates a thymidylate synthase (TS) reporter enzyme, which allows the E. coli strain D1210DthyA expressing this protein to grow in media lacking thymine. This sensor is capable of differentiating agonist compounds from antagonist compounds and has shown utility in the identification and characterization of a number of compounds that bind to human and animal estrogen receptors (ERs; Gawrys et al 2009, Gierach et al 2013. More recently, this strategy has been validated for subtype-selective human TR ligands using a small library of known selective ligands (Gierach et al 2012).…”

Section: Introductionmentioning

confidence: 99%

Analysis of the roles of mutations in thyroid hormone receptor-β by a bacterial biosensor system

Shi¹,

Meng²,

Wood³

2013

Journal of Molecular Endocrinology

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Mutations in thyroid hormone receptors (TRs) often lead to metabolic and developmental disorders, but patients with these mutations are difficult to treat with existing thyromimetic drugs. In this study, we analyzed six clinically observed mutations in the ligand-binding domain of the human TRb using an engineered bacterial hormone biosensor. Six agonist compounds, including triiodothyronine (T 3 ), thyroxine (T 4 ), 3,5,3 0 -triiodothyroacetic acid (Triac), GC-1, KB-141, and CO-23, and the antagonist NH-3 were examined for their ability to bind to each of the TRb mutants. The results indicate that some mutations lead to the loss of ability to bind to native ligands, ranging from several fold to several hundred fold, while other mutations completely abolish the ability to bind to any ligand. Notably, the effect of each ligand on each TRb mutant in this bacterial system is highly dependent on both the mutation and the ligand; some ligands were bound well by a wide variety of mutants, while other ligands lost their affinity for all but the WT receptor. This study demonstrates the ability of our bacterial system to differentiate agonist compounds from antagonist compounds and shows that one of the TRb mutations leads to an unexpected increase in antagonist ability relative to other mutations. These results indicate that this bacterial sensor can be used to rapidly determine ligand-binding ability and character for clinically relevant TRb mutants. Key Words" thyroid hormone receptor " ligand-binding assay " hormone receptor mutations " engineered biosensor

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Bacterial biosensors for evaluating potential impacts of estrogenic endocrine disrupting compounds in multiple species

Cited by 15 publications

References 58 publications

Intein Applications: From Protein Purification and Labeling to Metabolic Control Methods

Intein Applications: From Protein Purification and Labeling to Metabolic Control Methods

Environmental Application of Reporter-Genes Based Biosensors for Chemical Contamination Screening

Analysis of the roles of mutations in thyroid hormone receptor-β by a bacterial biosensor system

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