Insight into the biochemical mechanism of DNA helicases provided by bulk-phase and single-molecule assays

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2 Scopus citations

Abstract

There are multiple assays available that can provide insight into the biochemical mechanism of DNA helicases. For the first 22 years since their discovery, bulk-phase assays were used. These include gel-based, spectrophotometric, and spectrofluorometric assays that revealed many facets of these enzymes. From 2001, single-molecule studies have contributed additional insight into these DNA nanomachines to reveal details on energy coupling, step size, processivity as well as unique aspects of individual enzyme behavior that were masked in the averaging inherent in ensemble studies. In this review, important aspects of the study of helicases are discussed including beginning with active, nuclease-free enzyme, followed by several bulk-phase approaches that have been developed and still find widespread use today. Finally, two single-molecule approaches are discussed, and the resulting findings are related to the results obtained in bulk-phase studies.

Original languageEnglish (US)
Pages (from-to)348-360
Number of pages13
JournalMethods
Volume204
DOIs
StatePublished - Aug 2022

Keywords

  • DNA helicase
  • Magnetic tweezers
  • Optical trap
  • Pif1
  • RecG
  • SMARCAL1

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

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