Peptide fragments of DNA topoisomerase II with helix-forming and coiled-coil-forming properties act as inhibitors of the enzyme

Abstract

We have previously shown that a synthetic peptide (dL) consisting of amino acids 1013-1056 of human a topoisomerase II adopted an α-helix structure and formed a stable dimer coiled-coil in solution [Frere, V., Sourgen, F, Monnot, M., Troalen, E and Fermandjian, S. (1995) J. Biol. Chem. 270, 17502-17 507]. Here we studied two peptides, dP and dL short, which are related to dL but which have a double substitution Leu1026→Pro, Leu1037→Pro and a deletion of the 15 C-terminal residues, respectively. The peptides were studied for their ability to form α-helix structures, coiled coils, and to inhibit topoisomerase II activity. In combining circular dichroism spectra with AGADIR prediction for helix structures, we demonstrated that the dL short peptide, like its parent dL peptide, adopts an α-helix structure and can autoassociate into coiled-coils, while dP is completely devoid of such properties. Remarkably, only the dL and dL short peptides act as good inhibitors of topoisomerase II in various in vitro assays. However, the dL short peptide has a stronger helix potential and behaves as a much more potent inhibitor (5 μM versus 200 μM) compared to the dL peptide. All these data strongly suggest that the greater inhibitory effect demonstrated by the dL short peptide is related to its higher ability to form a stable amphiphilic helix, which in turn better recognizes its homologous helical segment in topoisomerase II. Finally, we propose that the dL and the dL short peptides could interfere with the enzymatic activity of topoisomersase II in modifying its autoassociation or translocation properties. Such peptides may serve as useful models for developing simpler and more specific inhibitors of topoisomerase II.