DNA targeting by metallopeptides: A comparison to peptide -derived natural products

Mark Anthony Lewis, Purdue University

Abstract

The DNA targeting of M(II)·Gly1-Gly2-His-derived metallopeptides was investigated via several methodologies. The high-throughput fluorescent intercalator displacement assay (HT-FID) was employed to investigate the DNA sequence-selective binding of the metallopeptides and peptide-derived natural products. The FID protocol was amenable to the investigation of natural products that exhibited DNA binding affinities greater than 1 x 10 3 M-1. In contrast, the FID protocol was not sensitive enough to elucidate the site-selective DNA binding exhibited by our M(II)·Gly 1-Gly2-His-derived metallopeptides. The metallopeptide-DNA interaction was also investigated via isothermal titration calorimetry (ITC) and surface plasmon resonance (SPR) two probeless techniques. The metallopeptides did not produce an ITC-detectible change in heat upon their interaction with DNA, and SPR experiments suggested that their DNA-association was weak and most likely the result of an electrostatic interaction. M(II)·Gly 1-Gly2-His-derived metallopeptides do not exhibit the equilibrium DNA-binding characteristics of netropsin or Co(III)·bleomycin, instead these are kinetically driven DNA damaging agents.

Degree

Ph.D.

Advisors

Long, Purdue University.

Subject Area

Biochemistry

Off-Campus Purdue Users:
To access this dissertation, please log in to our
proxy server
.

Share

COinS