Literature DB >> 27269812

A Platform to Enable the Pharmacological Profiling of Small Molecules in Gel-Based Electrophoretic Mobility Shift Assays.

Timothy L Foley1,2, Dorjbal Dorjsuren1, Thomas S Dexheimer1,3, Michael D Burkart4, William C Wight5, Anton Simeonov1.   

Abstract

We describe a polyacrylamide gel casting cassette that overcomes limitations of commercially available gel electrophoresis equipment. This apparatus molds a single polyacrylamide gel that can evaluate more than 200 samples in parallel, is loaded with a multichannel pipettor, and is flexible with respect to composition of the separating matrix. We demonstrate its use to characterize inhibitors of enzymes that modify protein and nucleic acid substrates. Throughputs of greater than 1000 samples per day were achieved when this system was paired with a quantitative laser-based imaging system, yielding data of remarkable quality.

Entities:  

Keywords:  electrophoretic mobility shift assay (EMSA); electrophoretic techniques; enzymatic assays and analyses; high-throughput screening (HTS); polyacrylamide gel electrophoresis (PAGE); small-molecule testing

Mesh:

Substances:

Year:  2016        PMID: 27269812      PMCID: PMC5269537          DOI: 10.1177/1087057116652895

Source DB:  PubMed          Journal:  J Biomol Screen        ISSN: 1087-0571


  11 in total

Review 1.  Experimental strategies for the identification of DNA-binding proteins.

Authors:  Michael F Carey; Craig L Peterson; Stephen T Smale
Journal:  Cold Spring Harb Protoc       Date:  2012-01-01

Review 2.  High-throughput screening assays for the identification of chemical probes.

Authors:  James Inglese; Ronald L Johnson; Anton Simeonov; Menghang Xia; Wei Zheng; Christopher P Austin; Douglas S Auld
Journal:  Nat Chem Biol       Date:  2007-08       Impact factor: 15.040

3.  Compound Management for Quantitative High-Throughput Screening.

Authors:  Adam Yasgar; Paul Shinn; Ajit Jadhav; Douglas Auld; Sam Michael; Wei Zheng; Christopher P Austin; James Inglese; Anton Simeonov
Journal:  JALA Charlottesv Va       Date:  2008-04

Review 4.  Novel trends in high-throughput screening.

Authors:  Lorenz M Mayr; Dejan Bojanic
Journal:  Curr Opin Pharmacol       Date:  2009-09-21       Impact factor: 5.547

Review 5.  Academic drug discovery: current status and prospects.

Authors:  Jeremy R Everett
Journal:  Expert Opin Drug Discov       Date:  2015-06-19       Impact factor: 6.098

6.  Synthesis and SAR studies of 5-(pyridin-4-yl)-1,3,4-thiadiazol-2-amine derivatives as potent inhibitors of Bloom helicase.

Authors:  Andrew S Rosenthal; Thomas S Dexheimer; Opher Gileadi; Giang H Nguyen; Wai Kit Chu; Ian D Hickson; Ajit Jadhav; Anton Simeonov; David J Maloney
Journal:  Bioorg Med Chem Lett       Date:  2013-08-13       Impact factor: 2.823

7.  Selective and cell-active inhibitors of the USP1/ UAF1 deubiquitinase complex reverse cisplatin resistance in non-small cell lung cancer cells.

Authors:  Junjun Chen; Thomas S Dexheimer; Yongxing Ai; Qin Liang; Mark A Villamil; James Inglese; David J Maloney; Ajit Jadhav; Anton Simeonov; Zhihao Zhuang
Journal:  Chem Biol       Date:  2011-11-23

8.  A strategy to discover inhibitors of Bacillus subtilis surfactin-type phosphopantetheinyl transferase.

Authors:  Adam Yasgar; Timothy L Foley; Ajit Jadhav; James Inglese; Michael D Burkart; Anton Simeonov
Journal:  Mol Biosyst       Date:  2009-10-13

9.  4-(3-Chloro-5-(trifluoromethyl)pyridin-2-yl)-N-(4-methoxypyridin-2-yl)piperazine-1-carbothioamide (ML267), a potent inhibitor of bacterial phosphopantetheinyl transferase that attenuates secondary metabolism and thwarts bacterial growth.

Authors:  Timothy L Foley; Ganesha Rai; Adam Yasgar; Thomas Daniel; Heather L Baker; Matias Attene-Ramos; Nicolas M Kosa; William Leister; Michael D Burkart; Ajit Jadhav; Anton Simeonov; David J Maloney
Journal:  J Med Chem       Date:  2014-01-22       Impact factor: 7.446

10.  Identification and characterization of inhibitors of human apurinic/apyrimidinic endonuclease APE1.

Authors:  Anton Simeonov; Avanti Kulkarni; Dorjbal Dorjsuren; Ajit Jadhav; Min Shen; Daniel R McNeill; Christopher P Austin; David M Wilson
Journal:  PLoS One       Date:  2009-06-01       Impact factor: 3.240

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  2 in total

1.  Horizontal Gel Electrophoresis for Enhanced Detection of Protein-RNA Complexes.

Authors:  Megan E Dowdle; Susanne Blaser Imboden; Sookhee Park; Sean P Ryder; Michael D Sheets
Journal:  J Vis Exp       Date:  2017-07-28       Impact factor: 1.355

2.  Discovering de novo peptide substrates for enzymes using machine learning.

Authors:  Lorillee Tallorin; JiaLei Wang; Woojoo E Kim; Swagat Sahu; Nicolas M Kosa; Pu Yang; Matthew Thompson; Michael K Gilson; Peter I Frazier; Michael D Burkart; Nathan C Gianneschi
Journal:  Nat Commun       Date:  2018-12-07       Impact factor: 14.919

  2 in total

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