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.com
Volume 10, Issue 8 (Suppl)
J Proteomics Bioinform, an open access journal
ISSN: 0974-276X
Structural Biology 2017
September 18-20, 2017
9
th
International Conference on
Structural Biology
September 18-20, 2017 Zurich, Switzerland
Cellular and biophysical pipeline for peroxisome proliferator-activated receptor (PPAR) delta agonist
screening
Natalia Bernardi Videira
1,2
, Artur Torres Cordeiro
1
and
Ana Carolina Migliorini Figueira
1
1
Center of Research in Energy and Materials, Brazil
2
University of Campinas, Brazil
P
eroxisome Proliferator Activated Receptors delta (PPARδ) has been associated with pathophysiological processes, such as
inflammation, obesity, dyslipidemia, diabetes, cancer, and cardiovascular diseases, being considered as new therapeutic
targets for these processes. Here, we developed and set up one way to perform a screening to drive PPARδ agonists. We use
methodologies capable of identify new molecules from compound libraries, which may work as this receptor’s ligand. The
first step in this screening pipeline is a valid cellular transactivation assay, as the primary search for potential compounds. We
developed one assay based on a cellular transactivation reporter gene technology, performed on a 96-well microplate with
support of automated pipette. The applied validation methodology was a combination of a thermal shift assay, used to check if
the compounds or extract components selected in the transactivation assay stabilize PPARδ tertiary structure; coupled with a
ANS quenching assay, which checks if the compound binds to the hydrophobic ligand binding pocket of PPARδ. Furthermore,
the quality of the cellular high-throughput screening (HTS) in stability and reliability was evaluated by the Z-factor, and a
natural extract library was used to validate the developed method. The results suggested that we developed a pipeline capable
to search compounds or extracts feasible and robust enough to measure PPARδ activation, tertiary structure stabilization and
ligand binding. As example, we could find one plant extract that contains interesting molecules, capable to binding and activate
PPARδ. In conclusion, this pipeline presented more efficacy in comparison to the single activation screening, because it can
exclude false-positives that may promote indirect PPARδ activation, without physical interaction with the receptor. Finally, this
approach may improve the effectiveness of screening agonists targeting PPARδ for drug development.
Biography
Natalia Bernardi Videira is a PhD student from Brazilian Biosciences National Laboratory (LNBio). LNBio is a laboratory dedicated to cutting-edge research and
innovation focused on biotechnology and drugs development. Her PhD project deals with the development of a screening pipeline for PPAR delta agonists. She
is currently in Switzerland for a 1-year research internship in the Center of Integrative Genomics, University of Lausanne. There she will study PPAR-dependent
regulations of skin cell responses to environmental insults under supervision of Dr. Michalik.
nataliabvideira@gmail.comNatalia Bernardi Videira et al., J Proteomics Bioinform 2017, 10:8(Suppl)
DOI: 10.4172/0974-276X-C1-0101