A simple DNA handle attachment method for single molecule mechanical manipulation experiments

Duyoung Min, Mark A. Arbing, Robert E. Jefferson, James U. Bowie*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

34 Citations (Scopus)

Abstract

Manipulating single molecules and systems of molecules with mechanical force is a powerful technique to examine their physical properties. Applying force requires attachment of the target molecule to larger objects using some sort of molecular tether, such as a strand of DNA. DNA handle attachment often requires difficult manipulations of the target molecule, which can preclude attachment to unstable, hard to obtain, and/or large, complex targets. Here we describe a method for covalent DNA handle attachment to proteins that simply requires the addition of a preprepared reagent to the protein and a short incubation. The handle attachment method developed here provides a facile approach for studying the biomechanics of biological systems.

Original languageEnglish
Pages (from-to)1535-1544
Number of pages10
JournalProtein Science
DOIs
Publication statusPublished - 1 Aug 2016

Keywords

  • forced unfolding
  • magnetic tweezers
  • membrane protein
  • optical tweezers
  • protein folding
  • SpyCatcher
  • SpyTag

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