nature11在G四方体和小分子之间建立A single-molecule platform for investigation.pdfVIP

ARTICLES PUBLISHED ONLINE: 28 AUGUST 2011 | DOI: 10.1038/NCHEM.1126 A single-molecule platform for investigation of interactions between G-quadruplexes and small-molecule ligands 1 1 2 3 ¨ 2 Deepak Koirala , Soma Dhakal , Beth Ashbridge , Yuta Sannohe , Raphael Rodriguez , Hiroshi Sugiyama3,4,5, Shankar Balasubramanian2,6,7 and Hanbin Mao1 * Ligands that stabilize the formation of telomeric DNA G-quadruplexes have potential as cancer treatments, because the G-quadruplex structure cannot be extended by telomerase, an enzyme over-expressed in many cancer cells. Understanding the kinetic, thermodynamic and mechanical properties of small-molecule binding to these structures is therefore important, but classical ensemble assays are unable to measure these simultaneously. Here, we have used a laser tweezers method to investigate such interactions. With a force jump approach, we observe that pyridostatin promotes the folding of telomeric G-quadruplexes. The increased mechanical stability of pyridostatin-bound G-quadruplex permits the determination of a dissociation constant Kd of 490 +++++80 nM. The free-energy change of binding obtained from a Hess-like process provides an identical Kd for pyridostatin and a Kd of 42 +++++3 mM for a weaker ligand RR110. We anticipate that this single-molecule platform can provide detailed insights into the mechanical, kinetic and thermodynamic properties of liganded bio-macromolecules, which have biological relevance. -quadruplex