title: Direct quantification of loop interaction and π-π stacking for G-quadruplex stability at the submolecular level.
abstract: The well-demonstrated biological functions of DNA G-quadruplex inside cells call for small molecules that can modulate these activities by interacting with G-quadruplexes. However, the paucity of the understanding of the G-quadruplex stability contributed from submolecular elements, such as loops and tetraguanine (G) planes (or G-quartets), has hindered the development of small-molecule binders. Assisted by click chemistry, herein, we attached pulling handles via two modified guanines in each of the three G-quartets in human telomeric G-quadruplex. Mechanical unfolding using these handles revealed that the loop interaction contributed more to the G-quadruplex stability than the stacking of G-quartets. This result was further confirmed by the binding of stacking ligands, such as telomestatin derivatives, which led to similar mechanical stability for all three G-quartets by significant reduction of loop interactions for the top and bottom G-quartets. The direct comparison of loop interaction and G-quartet stacking in G-quadruplex provides unprecedented insights for the design of more efficient G-quadruplex-interacting molecules. Compared to traditional experiments, in which mutations are employed to elucidate the roles of specific residues in a biological molecule, our submolecular dissection offers a complementary approach to evaluate individual domains inside a molecule with fewer disturbances to the native structure.
authors.affiliation: Department of Chemistry and Biochemistry and School of Biomedical Sciences, Kent State University , Kent, Ohio 44242, United States.
authors.forename: Chiran
authors.initials: C
authors.lastname: Ghimire
authors.forename: Soyoung
authors.initials: S
authors.lastname: Park
authors.forename: Keisuke
authors.initials: K
authors.lastname: Iida
authors.forename: Philip
authors.initials: P
authors.lastname: Yangyuoru
authors.forename: Haruka
authors.initials: H
authors.lastname: Otomo
authors.forename: Zhongbo
authors.initials: Z
authors.lastname: Yu
authors.forename: Kazuo
authors.initials: K
authors.lastname: Nagasawa
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Sugiyama
authors.forename: Hanbin
authors.initials: H
authors.lastname: Mao
mesh_terms: D002942:Circular Dichroism; D054856:G-Quadruplexes; D020349:Surface Plasmon Resonance