Characterization of DNA bound cyclic GMP-AMP synthase using atomic force microscopy imaging

Alexander Lushnikov, Richard Hooy, Jungsan Sohn, Alexey Krasnoslobodtsev

Research output: Chapter in Book/Report/Conference proceeding › Chapter

Abstract

The protocol described herein allows for acquiring topography images of DNA-protein complexes using Atomic Force Microscopy imaging. Since the very beginning of this method, AFM has been an indispensable tool for characterization of biomolecular complexes with exceptional capability of observing single complexes. This method can visualize structural characteristics of DNA-protein assemblies and evaluate differences between individual complexes. Although this protocol is generally applicable to a large number of various proteins complexed with DNA, we use cyclic G/AMP synthase (cGAS) enzyme as a case study for the protocol description.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
EditorsJungsan Sohn
PublisherAcademic Press Inc.
Pages157-166
Number of pages10
ISBN (Print)9780128183595
DOIs
StatePublished - 2019

Publication series

NameMethods in Enzymology
Volume625
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • Atomic force microscopy
  • Cyclic G/AMP synthase
  • DNA–protein complex
  • Mica surface modification
  • Topography imaging

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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    Lushnikov, A., Hooy, R., Sohn, J., & Krasnoslobodtsev, A. (2019). Characterization of DNA bound cyclic GMP-AMP synthase using atomic force microscopy imaging. In J. Sohn (Ed.), Methods in Enzymology (pp. 157-166). (Methods in Enzymology; Vol. 625). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2019.07.031