Step 2. Assay Development and Screening

  1. Natarajan A, Moerke N, Fan YH, Chen H, Christ WJ, Wagner G, Halperin JA. Synthesis of fluorescein labeled 7-methylguanosinemonophosphate. Bioorg Med Chem Lett. 2004 May 17;14(10):2657-60. doi: 10.1016/j.bmcl.2004.02.090. PMID: 15109672.

  2. Lokesh GL, Rachamallu A, Kumar GD, Natarajan A. High-throughput fluorescence polarization assay to identify small molecule inhibitors of BRCT domains of breast cancer gene 1. Anal Biochem. 2006 May 1;352(1):135-41. doi: 10.1016/j.ab.2006.01.025. Epub 2006 Feb 3. PMID: 16500609.

  3. Simeonov A, Yasgar A, Jadhav A, Lokesh GL, Klumpp C, Michael S, Austin CP, Natarajan A, Inglese J. Dual-fluorophore quantitative high-throughput screen for inhibitors of BRCT-phosphoprotein interaction. Anal Biochem. 2008 Apr 1;375(1):60-70. doi: 10.1016/j.ab.2007.11.039. Epub 2007 Dec 5. PMID: 18158907; PMCID: PMC3389998.

  4. Kumar EA, Charvet CD, Lokesh GL, Natarajan A. High-throughput fluorescence polarization assay to identify inhibitors of Cbl(TKB)-protein tyrosine kinase interactions. Anal Biochem. 2011 Apr 15;411(2):254-60. doi: 10.1016/j.ab.2010.11.038. Epub 2010 Dec 1. PMID: 21129358; PMCID: PMC3078584.

  5. Arnst JL, Davies CW, Raja SM, Das C, Natarajan A. High-throughput compatible fluorescence resonance energy transfer-based assay to identify small molecule inhibitors of AMSH deubiquitinase activity. Anal Biochem. 2013 Sep 1;440(1):71-7. doi: 10.1016/j.ab.2013.05.017. Epub 2013 Jun 5. PMID: 23747283; PMCID: PMC3784021.

  6. Al-Mugotir M, Kolar C, Vance K, Kelly DL, Natarajan A, Borgstahl GEO. A simple fluorescent assay for the discovery of protein-protein interaction inhibitors. Anal Biochem. 2019 Mar 15;569:46-52. doi: 10.1016/j.ab.2019.01.010. Epub 2019 Jan 30. PMID: 30707898; PMCID: PMC7968734.

3. Hit Identification