Cellular and Biophysical Pipeline for the Screening of Peroxisome Proliferator-Activated Receptor BetaDelta Agonists: Avoiding False Positives
Joint Authors
Videira, Natália Bernardi
Batista, Fernanda Aparecida Heleno
Torres Cordeiro, Artur
Figueira, Ana Carolina Migliorini
Source
Issue
Vol. 2018, Issue 2018 (31 Dec. 2018), pp.1-14, 14 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2018-04-12
Country of Publication
Egypt
No. of Pages
14
Main Subjects
Abstract EN
Peroxisome proliferator-activated receptor beta/delta (PPARß/δ) is considered a therapeutic target for metabolic disorders, cancer, and cardiovascular diseases.
Here, we developed one pipeline for the screening of PPARß/δ agonists, which reduces the cost, time, and false-positive hits.
The first step is an optimized 3-day long cellular transactivation assay based on reporter-gene technology, which is supported by automated liquid-handlers.
This primary screening is followed by a confirmatory transactivation assay and by two biophysical validation methods (thermal shift assay (TSA) and (ANS) fluorescence quenching), which allow the calculation of the affinity constant, giving more information about the selected hits.
All of the assays were validated using well-known commercial agonists providing trustworthy data.
Furthermore, to validate and test this pipeline, we screened a natural extract library (560 extracts), and we found one plant extract that might be interesting for PPARß/δ modulation.
In conclusion, our results suggested that we developed a cheaper and more robust pipeline that goes beyond the single activation screening, as it also evaluates PPARß/δ tertiary structure stabilization and the ligand affinity constant, selecting only molecules that directly bind to the receptor.
Moreover, this approach might improve the effectiveness of the screening for agonists that target PPARß/δ for drug development.
American Psychological Association (APA)
Videira, Natália Bernardi& Batista, Fernanda Aparecida Heleno& Torres Cordeiro, Artur& Figueira, Ana Carolina Migliorini. 2018. Cellular and Biophysical Pipeline for the Screening of Peroxisome Proliferator-Activated Receptor BetaDelta Agonists: Avoiding False Positives. PPAR Research،Vol. 2018, no. 2018, pp.1-14.
https://search.emarefa.net/detail/BIM-1212514
Modern Language Association (MLA)
Videira, Natália Bernardi…[et al.]. Cellular and Biophysical Pipeline for the Screening of Peroxisome Proliferator-Activated Receptor BetaDelta Agonists: Avoiding False Positives. PPAR Research No. 2018 (2018), pp.1-14.
https://search.emarefa.net/detail/BIM-1212514
American Medical Association (AMA)
Videira, Natália Bernardi& Batista, Fernanda Aparecida Heleno& Torres Cordeiro, Artur& Figueira, Ana Carolina Migliorini. Cellular and Biophysical Pipeline for the Screening of Peroxisome Proliferator-Activated Receptor BetaDelta Agonists: Avoiding False Positives. PPAR Research. 2018. Vol. 2018, no. 2018, pp.1-14.
https://search.emarefa.net/detail/BIM-1212514
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1212514