Cantilever Sensor for Nanomechanical Detection of Specific Protein Conformations

Mukhopadhyay, R.; Sumbayev, Vadim V.; Lorentzen, Martin; Kjems, Jørgen; Andreasen, Peter A.; Besenbacher, Flemming

doi:10.1021/nl051449z

Cited by 118 publications

(80 citation statements)

References 10 publications

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“…Each conformation is recognised by a unique combination of peptides, defining a specific peptide recognition pattern. We characterised ER and ER corformations induced by suspected environmental endocrine disruptors among pesticides (Sumbayev et al, 2005) and tested various nanotechnological assays based on peptide-receptor binding (Mukhopadyay et al, 2005;Hansen et al, 2007). Based on these data, it seems that peptide recognition patterns may serve as sensors of ER conformations.…”

Section: Page 4 Of 30mentioning

confidence: 99%

“…Brown (Boston, MA, USA) (Halachmi et al, 1990), using the QuickChange Site-Directed Mutagenesis Kit (Sumbayev et al, 2005). The introduction of the mutation was validated by DNA sequencing as described above.…”

Section: Site-directed Mutagenesis and Purification Of The Gst-er-lbmentioning

confidence: 99%

“…The bound fusion phage was detected with a HRP-conjugated anti-M13 monoclonal antibody and a peroxidase reaction (orthophenylenediamine/H 2 O 2 , Kem-En-Tek Diagnostics, Copenhagen, Denmark). Colour development was measured by a microplate reader as absorbance at 492 nm (Sumbayev et al, 2005).…”

Section: Phage Elisa With Full Length Ers and With Gst-er-lbdmentioning

confidence: 99%

“…All experiments were performed at 25 o C in a binding buffer of 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1 mM EDTA, 0.005% Tween 20, 1 mM sodium ascorbate (Mukhopadhyay et al, 2005). Peptides fused to the D1D2 protein were immobilized on CM5 chips to a density of 3526  612 RU per mm 2 (3 different experiments per peptide), using the standard…”

Section: Spr Analysesmentioning

confidence: 99%

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Novel modes of oestrogen receptor agonism and antagonism by hydroxylated and chlorinated biphenyls, revealed by conformation-specific peptide recognition patterns

Sumbayev

Jensen

Hansen

et al. 2008

Molecular and Cellular Endocrinology

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Please cite this article as: Sumbayev, V.V., Jensen, J.K., Hansen, J.A., Andreasen, P.A., Novel modes of oestrogen receptor agonism and antagonism by hydroxylated and chlorinated biphenyls, revealed by conformation-specific peptide recognition patterns, Molecular and Cellular Endocrinology (2007), doi:10.1016/j.mce.2008 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. defined by X-ray crystal structure analysis of receptors in complex with the side chain-less 5,11-cis-diethyl-5,6,11,12-tetrahydrochrysene-2,8-diol (THC). We have now used the ability of peptides selected from phagedisplayed peptide libraries to bind conformation-specifically to oestrogen receptor- and - LBDs to analyse conformations induced by THC and a group of chlorinated biphenyls and their aryl-hydroxylated metabolites, suspected of being environmental chemical disruptors. In oestrogen receptor- THC defined a "passive antagonist" peptide recognition pattern, which was also induced by several antagonistic hydroxylated biphenyls, while a clearly different peptide recognition pattern was induced by their chlorinated agonistic counterparts. In oestrogen receptor-, THC induced a conformation similar to that induced by oestriol and other oestrogen receptor- agonists, which, as evaluated by site-directed mutagenesis, have a functionally important interaction with oestrogen receptor- residue His524. We conclude that the peptide recognition pattern can be used to used classify suspected environmental endocrine disruptors according the oestrogen receptor-and -conformations they induce.

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Section: Page 4 Of 30mentioning

confidence: 99%

Section: Site-directed Mutagenesis and Purification Of The Gst-er-lbmentioning

confidence: 99%

Section: Phage Elisa With Full Length Ers and With Gst-er-lbdmentioning

confidence: 99%

Section: Spr Analysesmentioning

confidence: 99%

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Novel modes of oestrogen receptor agonism and antagonism by hydroxylated and chlorinated biphenyls, revealed by conformation-specific peptide recognition patterns

Sumbayev

Jensen

Hansen

et al. 2008

Molecular and Cellular Endocrinology

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“…They also provided a strong correlation between ligand induced ER conformations and their biological activities. 25 Previous studies using piezoresistive cantilevers, 26 electrochemical sensor, 27 and reflectometric interference spectroscopy 28 have successfully detected the ligand induced ER conformation change through specific peptide-probes.…”

Section: ■ Introductionmentioning

confidence: 99%

Assessment of Estrogenic Activity of Perfluoroalkyl Acids Based on Ligand-induced Conformation State of Human Estrogen Receptor

Gao

Guo

2012

Environ. Sci. Technol.

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Perfluoroalkyl acids (PFAAs) have been reported to interfere with the endocrine system in vivo by mimicking endogenous hormone activities and causing adverse effects. Some exoestrogens bind to estrogen receptor (ER) and subsequently induce an ERmediated response. The transcriptional activity of ER is regulated by its distinct conformational states that are the results of ligand binding. In this work, a biosensor based on surface plasmon resonance (SPR) technique was developed which can discriminate between agonist and antagonist of human ERα (hERα) by monitoring the conformation state of the protein induced by ligand binding. The biosensor utilized the specific interaction between hERα and conformation-selective peptides. Six PFAAs with different chain lengths and acid groups were tested by the biosensor, and perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) were found to be ER agonists. Kinetic analyses of direct interaction between PFAAs and hERα by SPR revealed that PFOS and PFOA were both weak binders of ER with K D values of 2.19 and 107 μM respectively, whereas the other four PFAAs did not bind with ER. To understand the differences in ER binding affinity and estrogenic activity among the six PFAAs, molecular docking based on the crystal structure of hERα ligand binding domain was performed. PFOS and PFOA were efficiently docked with hERα and formed hydrogen bonds with Arg394 in a manner similar to estradiol. Overall, the two 8-carbon PFAAs were assessed as weak agonists of hERα and are of potential concern.

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Chemical and Biochemical Sensors, 1. Fundamentals

Bârsan

Gauglitz

Oprea

et al. 2016

Ullmann's Encyclopedia of Industrial Chemistry

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Cantilever Sensor for Nanomechanical Detection of Specific Protein Conformations

Cited by 118 publications

References 10 publications

Novel modes of oestrogen receptor agonism and antagonism by hydroxylated and chlorinated biphenyls, revealed by conformation-specific peptide recognition patterns

Novel modes of oestrogen receptor agonism and antagonism by hydroxylated and chlorinated biphenyls, revealed by conformation-specific peptide recognition patterns

Assessment of Estrogenic Activity of Perfluoroalkyl Acids Based on Ligand-induced Conformation State of Human Estrogen Receptor

Chemical and Biochemical Sensors, 1. Fundamentals

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