Topoisomerase II-drug interaction domains: Identification of substituents on etoposide that interact with the enzyme

Amy M. Wilstermann, Ryan P. Bender, Murrell Godfrey, Sungjo Choi, Clemens Anklin, David B Berkowitz, Neil Osheroff, David E. Graves

Research output: Contribution to journalArticle

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Abstract

Etoposide is one of the most successful chemotherapeutic agents used for the treatment of human cancers. The drug kills cells by inhibiting the ability of topoisomerase II to ligate nucleic acids that it cleaves during the double-stranded DNA passage reaction. Etoposide is composed of a polycyclic ring system (rings A-D), a glycosidic moiety at the C4 position, and a pendent ring (E-ring) at the C1 position. Although drug-enzyme contacts, as opposed to drug-DNA interactions, mediate the entry of etoposide into the topoisomerase II-drug-DNA complex, the substituents on etoposide that interact with the enzyme have not been identified. Therefore, saturation transfer difference [ 1H]-nuclear magnetic resonance spectroscopy and protein-drug competition binding assays were employed to define the groups on etoposide that associate with yeast topoisomerase II and human topoisomerase IIα. Results indicate that the geminal protons of the A-ring, the H5 and H8 protons of the B-ring, and the H2′ and H6′ protons and the 3′- and 5′-methoxyl protons of the pendent E-ring interact with both enzymes in the binary protein-ligand complexes. In contrast, no significant nuclear Overhauser enhancement signals arising from the C-ring, the D-ring, or the C4 glycosidic moiety were observed with either enzyme, suggesting that there is limited or no contact between these portions of etoposide and topoisomerase II in the binary complex. The functional importance of E-ring substituents was confirmed by topoisomerase II-mediated DNA cleavage assays.

Original languageEnglish (US)
Pages (from-to)8217-8225
Number of pages9
JournalBiochemistry
Volume46
Issue number28
DOIs
StatePublished - Jul 17 2007

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Drug interactions
Type II DNA Topoisomerase
Etoposide
Drug Interactions
Protons
Enzymes
Pharmaceutical Preparations
DNA
Assays
DNA Cleavage
Yeast
Nucleic Acids
Nuclear magnetic resonance spectroscopy
Proteins
Magnetic Resonance Spectroscopy
Yeasts
Ligands

ASJC Scopus subject areas

  • Biochemistry

Cite this

Topoisomerase II-drug interaction domains : Identification of substituents on etoposide that interact with the enzyme. / Wilstermann, Amy M.; Bender, Ryan P.; Godfrey, Murrell; Choi, Sungjo; Anklin, Clemens; Berkowitz, David B; Osheroff, Neil; Graves, David E.

In: Biochemistry, Vol. 46, No. 28, 17.07.2007, p. 8217-8225.

Research output: Contribution to journalArticle

Wilstermann, AM, Bender, RP, Godfrey, M, Choi, S, Anklin, C, Berkowitz, DB, Osheroff, N & Graves, DE 2007, 'Topoisomerase II-drug interaction domains: Identification of substituents on etoposide that interact with the enzyme', Biochemistry, vol. 46, no. 28, pp. 8217-8225. https://doi.org/10.1021/bi700272u
Wilstermann, Amy M. ; Bender, Ryan P. ; Godfrey, Murrell ; Choi, Sungjo ; Anklin, Clemens ; Berkowitz, David B ; Osheroff, Neil ; Graves, David E. / Topoisomerase II-drug interaction domains : Identification of substituents on etoposide that interact with the enzyme. In: Biochemistry. 2007 ; Vol. 46, No. 28. pp. 8217-8225.
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