Fluorescent biosensors to investigate helicase activity

Methods Mol Biol. 2010;587:13-27. doi: 10.1007/978-1-60327-355-8_2.

Abstract

ATP-driven translocation of helicases along DNA can be assayed in several ways. Reagentless biosensors, based on fluorophore-protein adducts, provide convenient ways for real-time assays of both the separation of dsDNA and the hydrolysis of ATP. Single-stranded DNA can be assayed using a modified single-stranded DNA-binding protein (SSB), and phosphate production during ATP hydrolysis can be measured by a modified phosphate-binding protein. Advantages and limitations of these approaches are compared with those of other types of measurements.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Assay / methods
  • Biosensing Techniques*
  • DNA Helicases / metabolism
  • DNA, Single-Stranded / metabolism*
  • DNA-Binding Proteins / metabolism
  • Fluorescent Dyes / metabolism*

Substances

  • DNA, Single-Stranded
  • DNA-Binding Proteins
  • Fluorescent Dyes
  • DNA Helicases