Fluorescent biosensors to investigate helicase activity.
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.
Webb
Martin R
MR
MRC National Institute for Medical Research, The Ridgeway, Mill Hill, London, UK.
D001681:Biological Assay / Q000379:methods; D015374:Biosensing Techniques; D004265:DNA Helicases / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism*
D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes; D004265:DNA Helicases