Biolayer interferometry provides a robust method for detecting DNA binding small molecules in microbial extracts
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RESEARCH PAPER
Biolayer interferometry provides a robust method for detecting DNA binding small molecules in microbial extracts Ross D. Overacker 1 & Birte Plitzko 1 & Sandra Loesgen 1,2 Received: 3 September 2020 / Revised: 24 October 2020 / Accepted: 17 November 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract DNA replication is an exceptional point of therapeutic intervention for many cancer types and several small molecules targeting DNA have been developed into clinically used antitumor agents. Many of these molecules are naturally occurring metabolites from plants and microorganisms, such as the widely used chemotherapeutic doxorubicin. While natural product sources contain a vast number of DNA binding small molecules, isolating and identifying these molecules is challenging. Typical screening campaigns utilize time-consuming bioactivity-guided fractionation approaches, which use sequential rounds of cell-based assays to guide the isolation of active compounds. In this study, we explore the use of biolayer interferometry (BLI) as a tool for rapidly screening natural product sources for DNA targeting small molecules. We first verified that BLI robustly detected DNA binding using designed GC- and AT-rich DNA oligonucleotides with known DNA intercalating, groove binding, and covalent binding agents including actinomycin D (1), doxorubicin (2), ethidium bromide (3), propidium iodide (4), Hoechst 33342 (5), and netropsin (6). Although binding varied with the properties of the oligonucleotides, measured binding affinities agreed with previously reported values. We next utilized BLI to screen over 100 bacterial extracts from our microbial library for DNA binding activity and found three highly active extracts. Binding-guided isolation was used to isolate the active principle component from each extract, which were identified as echinomycin (8), actinomycin V (9), and chartreusin (10). This biosensorbased DNA binding screen is a novel, low-cost, easy to use, and sensitive approach for medium-throughput screening of complex chemical libraries. Keywords DNA intercalation . DNA groove binding . Biolayer interferometry . Microbial natural products
Introduction DNA targeting drugs are known to show significant antibacterial [1], antiviral [2], and anticancer [3, 4] bioactivities. In particular, DNA binding chemotherapeutic agents take advantage of the fact that cancer cells are highly prone to DNA damage due to their lenient DNA repair capabilities and increased propagation resulting from their ability to bypass certain cell-cycle checkpoints [5, 6]. The first DNA binding chemotherapeutics were developed in the 1950s from
* Sandra Loesgen [email protected] 1
Department of Chemistry, Oregon State University, Corvallis, OR 97331, USA
2
Present address: Whitney Laboratory for Marine Bioscience, Department of Chemistry, University of Florida, St. Augustine, FL 32080, USA
anthracyclines isolated from a culture of Streptomyces purpurascens [7]. Originally, anthracyclines were pursu
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