The binding mode of the DNA bisintercalator luzopeptin investigated using atomic force microscopy.
The luzopeptins are DNA bisintercalating antibiotics that contain a decadepsipeptide to which are attached two quinoline chromophores. We have used atomic force microscopy (AFM) to investigate the interaction between luzopeptin B and DNA in an attempt to shed light on the binding mode of this antibiotic. AFM images provided contour lengths which were used as a direct measure of bisintercalation. Binding of luzopeptin B was investigated using two different DNA sequences, one having a GC content of 42% and the other 59%, which revealed a higher degree of bisintercalation into the DNA sequences having the lower GC content. The measured increment in contour length was found to plateau at values corresponding to binding of one drug molecule every 40 and 72 bp to the 42 and 59% GC sequences, respectively. In addition to the length increase, a higher proportion of DNA molecules displaying complex morphology was observed as the concentration of luzopeptin was increased. Such molecules were not included in the measurements of contour length. We propose that the various manifestations of complex morphology arise from both inter- and intramolecular cross-linking of the DNA caused by binding of luzopeptin, providing direct evidence of cross-linked species by AFM imaging.
Berge
Torunn
T
Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, UK. tb230@cam.ac.uk
Haken
E Lucy
EL
Waring
Michael J
MJ
Henderson
Robert M
RM
D001482:Base Composition; D001483:Base Sequence; D001665:Binding Sites; D003432:Cross-Linking Reagents / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D007091:Image Processing, Computer-Assisted; D007364:Intercalating Agents / Q000737:chemistry; D018625:Microscopy, Atomic Force / Q000379:methods*; D010456:Peptides, Cyclic / Q000737:chemistry*; D010957:Plasmids / Q000737:chemistry; D011804:Quinolines / Q000737:chemistry*; D047630:Depsipeptides
D003432:Cross-Linking Reagents; D004247:DNA; D007364:Intercalating Agents; D010456:Peptides, Cyclic; D011804:Quinolines; C029831:luzopeptins; D047630:Depsipeptides