Single-Molecule Imaging of Enzymatic Reactions on DNA Origami

  • An Yan
  • , Lele Sun
  • , Di Li*
  • *Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

The interaction between enzymes is very important for understanding of enzyme functions and shed light on enzymatic reaction mechanisms. With the development of DNA nanotechnology, DNA origami has become a powerful tool for analyzing the dynamic behavior of enzyme molecules. In this protocol, a method for imaging and analysis of single-molecule cascade enzyme reactions on DNA origami raft by total internal reflection fluorescence microscopy (TIRFM) is described. Through trajectory analysis and calculation, the diffusion of downstream enzymes in enzymatic reaction and chemotaxis of enzymatic reactions were elucidated at the single molecular level.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages131-145
Number of pages15
DOIs
StatePublished - 2023

Publication series

NameMethods in Molecular Biology
Volume2639
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • DNA origami
  • Enzyme cascades
  • Protein modification
  • Single molecular imaging
  • TIRFM

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