ArticleCombinatorial chemistry & high throughput screening2009
Development and validation of a high-density fluorescence polarization-based assay for the trypanosoma RNA triphosphatase TbCet1.
Article in Combinatorial chemistry & high throughput screening, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 12 citations in OpenAlex.
- High-throughput screening and structure-guided design of small molecules enable modulation of SAL-PAP stress signaling.Journal of experimental botany · 2026Article
- Enzymatic Assays to Explore Viral mRNA Capping Machinery.Chembiochem : a European journal of chemical biology · 2021Review
- Utility of Bioluminescent Homogeneous Nucleotide Detection Assays in Measuring Activities of Nucleotide-Sugar Dependent Glycosyltransferases and Studying Their Inhibitors.Molecules (Basel, Switzerland) · 2021Article
- Crystal structures of the RNA triphosphatase fromThe Journal of biological chemistry · 2020Article
- Article
- A 1536-well fluorescence polarization assay to screen for modulators of the MUSASHI family of RNA-binding proteins.Combinatorial chemistry & high throughput screening · 2014Article
- Comparison of luminescence ADP production assay and radiometric scintillation proximity assay for Cdc7 kinase.Combinatorial chemistry & high throughput screening · 2011Article
- Fluorescence polarization assays in small molecule screening.Expert opinion on drug discovery · 2011Article
- Curcumin decreases amyloid-beta peptide levels by attenuating the maturation of amyloid-beta precursor protein.The Journal of biological chemistry · 2010Article
- Development and validation of a transcreener assay for detection of AMP- and GMP-producing enzymes.Assay and drug development technologies · 2010Article
- Identification and preliminary characterization of novel small molecules that inhibit growth of human lung adenocarcinoma cells.Journal of biomolecular screening · 2009Article
- A miniaturized 1536-well format gamma-secretase assay.Assay and drug development technologies · 2009Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
RNA triphosphatases are attractive and mostly unexplored therapeutic targets for the development of broad spectrum antiprotozoal, antiviral and antifungal agents. The use of malachite green as a readout for phosphatases is well characterized and widely employed. However, the reaction depends on high quantities of inorganic phosphate to be generated, which makes this assay not easily amenable to screening in 1536-well format. The overly long reading times required also prohibit its use to screen large chemical libraries. To overcome these limitations, we sought to develop a fluorescence polarization (FP) -based assay for triphosphatases, compatible with miniaturization and fast readouts. For this purpose, we took advantage of the nucleoside triphosphatase activity of this class of enzyme to successfully adapt the Transcreener ADP assay based on the detection of generated ADP by immunocompetition fluorescence polarization to the RNA triphosphatase TbCet1 in 1536-well format. We also tested the performance of this newly developed assay in a pilot screen of 3,000 compounds and we confirmed the activity of the obtained hits. We present and discuss our findings and their importance for the discovery of novel drugs by high throughput screening.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.