+ Site Statistics
+ Search Articles
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ PDF Full Text
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Translate
+ Recently Requested

First identification of small-molecule inhibitors of Pontin by combining virtual screening and enzymatic assay

First identification of small-molecule inhibitors of Pontin by combining virtual screening and enzymatic assay

Biochemical Journal 443(2): 549-559

The human protein Pontin, which belongs to the AAA+ (ATPases associated with various cellular activities) family, is overexpressed in several cancers and its silencing in vitro leads to tumour cell growth arrest and apoptosis, making it a good target for cancer therapy. In particular, high levels of expression were found in hepatic tumours for which the therapeutic arsenal is rather limited. The three-dimensional structure of Pontin has been resolved previously, revealing a hexameric assembly with one ADP molecule co-crystallized in each subunit. Using Vina, DrugScore and Xscore, structure-based virtual screening of 2200 commercial molecules was conducted into the ATP-binding site formed by a dimer of Pontin in order to prioritize the best candidates. Complementary to the in silico screening, a versatile and sensitive colorimetric assay was set up to measure the disruption of the ATPase activity of Pontin. This assay allowed the determination of inhibition curves for more than 20 top-scoring compounds, resulting in the identification of four ligands presenting an inhibition constant in the micromolar concentration range. Three of them inhibited tumour cell proliferation. The association of virtual screening and experimental assay thus proved successful for the discovery of the first small-molecule inhibitors of Pontin.

(PDF emailed within 0-6 h: $19.90)

Accession: 053249653

Download citation: RISBibTeXText

PMID: 22273052

DOI: 10.1042/BJ20111779

Related references

Identification of novel DHFR inhibitors for treatment of tuberculosis by combining virtual screening with in vitro activity assay. Journal of Biomolecular Structure & Dynamics: 1-18, 2018

Identification of new GATA4-small molecule inhibitors by structure-based virtual screening. Bioorganic & Medicinal Chemistry 19(5): 1734-1742, 2011

Virtual screening based identification of novel small-molecule inhibitors targeted to the HIV-1 capsid. Bioorganic & Medicinal Chemistry 19(1): 77-90, 2011

Identification of Small Molecule Inhibitors of BCR/ABL Tyrosine Kinase through Structure Based Virtual Screening. Blood 100(11): Abstract No 1234, November 16, 2002

Characterization of small molecule binding. I. Accurate identification of strong inhibitors in virtual screening. Journal of Chemical Information and Modeling 53(1): 114-122, 2013

Application of Virtual Screening Approaches for the Identification of Small Molecule Inhibitors of the Methyllysine Reader Protein Spindlin1. Methods in Molecular Biology 1824: 347-370, 2018

A high throughput screening assay system for the identification of small molecule inhibitors of gsp. Plos One 9(3): E90766-E90766, 2015

Identification and characterization of small molecule inhibitors of signal transducer and activator of transcription 3 (STAT3) signaling pathway by virtual screening. Bioorganic & Medicinal Chemistry Letters 23(7): 2225-2229, 2013

Identification of High Affinity Non-Peptidic Small Molecule Inhibitors of MDM2-p53 Interactions through Structure-Based Virtual Screening Strategies. Asian Pacific Journal of Cancer Prevention 16(9): 3759-3765, 2016

Identification of Small-Molecule Inhibitors of the HuR/RNA Interaction Using a Fluorescence Polarization Screening Assay Followed by NMR Validation. Plos One 10(9): E0138780-E0138780, 2016

Frontal affinity chromatography with MS detection of EphB2 tyrosine kinase receptor. 2. Identification of small-molecule inhibitors via coupling with virtual screening. Journal of Medicinal Chemistry 48(9): 3221-3230, 2005

A fluorescence polarization based screening assay for identification of small molecule inhibitors of the PICK1 PDZ domain. Combinatorial Chemistry & High Throughput Screening 14(7): 590-600, 2014

High-throughput screening assay for identification of small molecule inhibitors of Aurora2/STK15 kinase. Journal of Biomolecular Screening 9(5): 391-397, 2004

Identification of novel small-molecule histone deacetylase inhibitors by medium-throughput screening using a fluorigenic assay. Biochemical Journal 413(1): 143-150, 2008

High-throughput screening AlphaScreen assay for identification of small-molecule inhibitors of ubiquitin E3 ligase SCFSkp2-Cks1. Journal of Biomolecular Screening 18(8): 910-920, 2014