Dissecting the Dynamic Pathways of Stereoselective DNA Threading Intercalation
Artikel i vetenskaplig tidskrift, 2016

DNA intercalators that have high affinity and slow kinetics are developed for potential DNA-targeted therapeutics. Although many natural intercalators contain multiple chiral subunits, only intercalators with a single chiral unit have been quantitatively probed. Dumbbell-shaped DNA threading intercalators represent the next order of structural complexity relative to simple intercalators, and can provide significant insights into the stereoselectivity of DNA-ligand intercalation. We investigated DNA threading intercalation by binuclear ruthenium complex [mu-dppzip(phen)(4)Ru-2](4+) (Piz). Four Piz stereoisomers are defined by the chirality of the intercalating subunit (Ru(phen)(2)dppz) and the distal subunit (Ru(phen)(2)ip), respectively, each of which can be either right-handed (Delta) or left-handed (Lambda). We used optical tweezers to measure single DNA molecule elongation due to threading intercalation, revealing force-dependent DNA intercalation rates and equilibrium dissociation constants. The force spectroscopy analysis provided the zero-force DNA binding affinity, the equilibrium DNA-ligand elongation Delta x(eq), and the dynamic DNA structural deformations during ligand association x(on) and dissociation x(off). We found that Piz stereoisomers exhibit over 20-fold differences in DNA binding affinity, from a K-d of 27 +/- 3 nM for (Delta,Lambda)-Piz to a K-d of 622 +/- 55 nM for (Lambda,Delta)-Piz. The striking affinity decrease is correlated with increasing Delta x(eq) from 0.30 +/- 0.02 to 0.48 +/- 0.02 nm and x(on) from 0.25 +/- 0.01 to 0.46 +/- 0.02 nm, but limited x(off) changes. Notably, the affinity and threading kinetics is 10-fold enhanced for right-handed intercalating subunits, and 2- to 5-fold enhanced for left-handed distal subunits. These findings demonstrate sterically dispersed transition pathways and robust DNA structural recognition of chiral intercalators, which are critical for optimizing DNA binding affinity and kinetics.

single dna

actinomycin-d

binuclear ruthenium complex

molecule

optical tweezers

binding-kinetics

noncooperative binding

equilibrium

affinity

force spectroscopy

Biophysics

Författare

A. A. Almaqwashi

Northeastern University

Johanna Andersson

Chalmers, Kemi och kemiteknik, Tillämpad kemi, Farmaceutisk teknologi

Per Lincoln

Chalmers, Kemi och kemiteknik, Kemi och biokemi, Fysikalisk kemi

I. Rouzina

Ohio State University

Fredrik Westerlund

Chalmers, Biologi och bioteknik, Kemisk biologi

M. C. Williams

Northeastern University

Biophysical Journal

0006-3495 (ISSN) 1542-0086 (eISSN)

Vol. 110 6 1255-1263

Styrkeområden

Nanovetenskap och nanoteknik

Livsvetenskaper och teknik

Ämneskategorier

Biofysik

DOI

10.1016/j.bpj.2016.02.016

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Senast uppdaterat

2018-04-09