利用纳米孔对DNA-蛋白质复合物进行单分子分析。

Single-molecule analysis of DNA-protein complexes using nanopores.

作者信息

Hornblower Breton, Coombs Amy, Whitaker Richard D, Kolomeisky Anatoly, Picone Stephen J, Meller Amit, Akeson Mark

机构信息

Department of Chemistry & Biomolecular Engineering, University of California, Santa Cruz, California 95064, USA.

出版信息

Nat Methods. 2007 Apr;4(4):315-7. doi: 10.1038/nmeth1021. Epub 2007 Mar 4.

DOI:10.1038/nmeth1021

PMID:17339846

Abstract

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.

摘要

我们提出了一种通过电压驱动单链DNA分子穿过蛋白质纳米孔来快速测量DNA-蛋白质相互作用的方法。施加于单个单链DNA-核酸外切酶I复合物的电力将两个分子拉开，同时离子电流探测复合物的解离速率。纳米孔力谱（NFS）揭示了影响复合物解离的能量屏障。该方法可使用蛋白质或固态纳米孔装置应用于其他核酸-蛋白质复合物。