Single-molecule analysis of DNA-protein complexes using nanopores

被引：241

作者：

Hornblower, Breton

Coombs, Amy

Whitaker, Richard D.

Kolomeisky, Anatoly

Picone, Stephen J.

Meller, Amit

Akeson, Mark

机构：

[1] Univ Calif Santa Cruz, Dept Chem, Santa Cruz, CA 95064 USA

[2] Univ Calif Santa Cruz, Dept Biomol Engn, Santa Cruz, CA 95064 USA

[3] Boston Univ, Dept Biomed & Phys, Boston, MA 02215 USA

[4] Boston Univ, Dept Phys, Boston, MA 02215 USA

[5] New England Biolabs Inc, Ipswich, MA 01938 USA

[6] Rice Univ, Dept Chem, Houston, TX 77005 USA

来源：

NATURE METHODS | 2007年 / 4卷 / 04期

关键词：

D O I：

10.1038/NMETH1021

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We present a method for rapid measurement of DNA-protein interactions using voltage-driven threading of single DNA molecules through a protein nanopore. Electrical force applied to individual ssDNA-exonuclease I complexes pulls the two molecules apart, while ion current probes the dissociation rate of the complex. Nanopore force spectroscopy (NFS) reveals energy barriers affecting complex dissociation. This method can be applied to other nucleic acid-protein complexes, using protein or solid-state nanopore devices.

引用

页码：315 / 317

页数：3

共 15 条

[1] Microsecond time-scale discrimination among polycytidylic acid, polyadenylic acid, and polyuridylic acid as homopolymers or as segments within single RNA molecules [J].