NMR studies on self‐complementary oligonucleotides conjugated with methylene blue

A carboxyl‐functionalized methylene blue (MB) derivative was synthesized and covalently coupled to three CG‐rich self‐complementary 2′‐deoxyoligonucleotides at their 5′‐end. Thermodynamic and structural details about the interactions between the dye and oligonucleotide duplexes were investigated employing ultraviolet (UV) melting and 1 H nuclear magnetic resonance (NMR) experiments. In contrast to previous findings on MB binding, no specific intercalation or binding in the minor or major groove of the double helix was found in a 100 m M NaCl buffer. Rather, proton chemical shift changes in the conjugates provide ample evidence for weak dye–DNA interactions largely through external MB stacking on the terminal base pairs. © 2005 Wiley Periodicals, Inc. Biopolymers 79: 335–343, 2005 This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com