FRET Assay for Ligands Targeting the Bacterial A-Site RNA.

Aminoglycosides / chemistry Coumarins / chemistry Fluorescence Fluorescence Resonance Energy Transfer / methods Ligands RNA, Bacterial / chemistry Uridine / chemistry
A-site binders Aminoglycosides Bacterial A-site FRET assay Fluorescence spectroscopy Fluorescent nucleosides

Journal

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969

Informations de publication

Date de publication:
2019
Historique:
entrez: 25 4 2019
pubmed: 25 4 2019
medline: 7 9 2019
Statut: ppublish

Résumé

A robust, fluorescence-based analysis and discovery platform is described for bacterial A-site binders. The assay relies on an incorporated isomorphic fluorescent uridine analog, which substitutes the A-site's U1406 and serves as a FRET donor to an A-site bound coumarin-labeled aminoglycoside that serves as the FRET acceptor. Binding efficiency of unlabeled A-site ligands can be determined by competition experiments, where the acceptor-labeled aminoglycoside is displaced. The replacement efficiency is gauged by the concentration-dependent loss of the sensitized FRET acceptor's signal with concomitant restoration of the donor's emission. Plotting the relative emission intensity of both the donor and acceptor as a function of ligand concentration followed by fitting of the data points to a dose-response curve yields IC

Identifiants

DOI: 10.1007/978-1-4939-9216-4_16 PMID: 31016707
pubmed: 31016707
doi: 10.1007/978-1-4939-9216-4_16
doi:

Substances chimiques

Aminoglycosides 0
Coumarins 0
Ligands 0
RNA, Bacterial 0
coumarin A4VZ22K1WT
Uridine WHI7HQ7H85

Types de publication

Journal Article Research Support, N.I.H., Extramural

Langues

eng

Sous-ensembles de citation

IM

Pagination

251-260

Auteurs

Renatus W Sinkeldam (RW)

Office of Technology Management, Washington University in St. Louis, St. Louis, MO, USA.

Yitzhak Tor (Y)

Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, USA. ytor@ucsd.edu.

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Classifications MeSH

questionsmedicales.fr › Glucides › Hétérosides › Aminosides › FRET Assay for Ligands Targeting the Bacterial...
questionsmedicales.fr › Composés hétérocycliques › Composés hétérocycliques à cycles fusionnés › Composés hétérobicycliques › Benzopyranes › Coumarines › FRET Assay for Ligands Targeting the Bacterial...
questionsmedicales.fr › Phénomènes physiques › Phénomènes optiques › Fluorescence › FRET Assay for Ligands Targeting the Bacterial...
questionsmedicales.fr › Phénomènes chimiques › Phénomènes biochimiques › Transfert d'énergie par résonance de fluorescence › FRET Assay for Ligands Targeting the Bacterial...
questionsmedicales.fr › Actions chimiques et utilisations › Utilisations spécialisées de produits chimiques › Produits chimiques de laboratoire › Ligands › FRET Assay for Ligands Targeting the Bacterial...
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ARN › ARN bactérien › FRET Assay for Ligands Targeting the Bacterial...
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Nucléosides › Ribonucléosides › Uridine › FRET Assay for Ligands Targeting the Bacterial...