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

Fluorescent biosensors to investigate helicase activity.

Author information

1
MRC National Institute for Medical Research, The Ridgeway, Mill Hill, London, UK.

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.

PMID:
20225139
DOI:
10.1007/978-1-60327-355-8_2
[Indexed for MEDLINE]

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