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Stefanovic B, Stefanovic L. Screening for antifibrotic compounds using high throughput system based on fluorescence polarization. Biology (Basel). 2014;3(2):281-94doi: 10.3390/biology3020281.
Stefanovic, B., & Stefanovic, L. (2014). Screening for antifibrotic compounds using high throughput system based on fluorescence polarization. Biology, 3(2), 281-94. https://doi.org/10.3390/biology3020281
Stefanovic, Branko, and Stefanovic, Lela. "Screening for antifibrotic compounds using high throughput system based on fluorescence polarization." Biology vol. 3,2 (2014): 281-94. doi: https://doi.org/10.3390/biology3020281
Stefanovic B, Stefanovic L. Screening for antifibrotic compounds using high throughput system based on fluorescence polarization. Biology (Basel). 2014 Apr 10;3(2):281-94. doi: 10.3390/biology3020281. PMID: 24833510; PMCID: PMC4085608.
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Biology (Basel). 2014 Apr 10;3(2):281-94. doi: 10.3390/biology3020281.

Screening for antifibrotic compounds using high throughput system based on fluorescence polarization.

Biology

Branko Stefanovic, Lela Stefanovic

Affiliations

  1. College of Medicine, Florida State University, 1115 W. Call St., Tallahassee, FL 32306, USA. [email protected].
  2. College of Medicine, Florida State University, 1115 W. Call St., Tallahassee, FL 32306, USA. [email protected].

PMID: 24833510 PMCID: PMC4085608 DOI: 10.3390/biology3020281

Abstract

Fibroproliferative diseases are one of the leading causes of death worldwide. They are characterized by reactive fibrosis caused by uncontrolled synthesis of type I collagen. There is no cure for fibrosis and development of therapeutics that can inhibit collagen synthesis is urgently needed. Collagen α1(I) mRNA and α2(I) mRNA encode for type I collagen and they have a unique 5' stem-loop structure in their 5' untranslated regions (5'SL). Collagen 5'SL binds protein LARP6 with high affinity and specificity. The interaction between LARP6 and the 5'SL is critical for biosynthesis of type I collagen and development of fibrosis in vivo. Therefore, this interaction represents is an ideal target to develop antifibrotic drugs. A high throughput system to screen for chemical compounds that can dissociate LARP6 from 5'SL has been developed. It is based on fluorescence polarization and can be adapted to screen for inhibitors of other protein-RNA interactions. Screening of 50,000 chemical compounds yielded a lead compound that can inhibit type I collagen synthesis at nanomolar concentrations. The development, characteristics, and critical appraisal of this assay are presented.

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