title: Unimolecular Chemically Modified DNA Fluorescent Probe for One-Step Quantitative Measurement of the Activity of Human Apurinic/Apyrimidinic Endonuclease 1 in Biological Samples.
abstract: A novel DNA structure containing a 3' internal-loop modified abasic site has been constructed which enables effective differentiation between apurinic/apyrimidinic endonuclease (APE1) and nonspecific endonuclease (DNase I). When this unique substrate structure is employed, a double-loop frayed-end chimeric fluorescent probe is successfully developed for quantitative measurement of the activity of APE1 in biological samples without the need of additional cleanup or preconcentration steps. The method is simple and rapid and has a single-step with a linear working range between 0.1 and 5.0 U/mL and a lower limit of detection of 0.1 U/mL. It holds great potential in real-time monitoring of the variation of intracellular and extracellular APE1, which will be very useful for further understanding of the DNA repair pathways in different organisms.
authors.affiliation: Beijing National Laboratory for Molecular Sciences, MOE Key Laboratory of Bioorganic Chemistry and Molecular Engineering, College of Chemistry and Molecular Engineering, Peking University , Beijing 100871, China.
authors.forename: Simin
authors.initials: S
authors.lastname: Fang
authors.affiliation: Beijing National Laboratory for Molecular Sciences, MOE Key Laboratory of Bioorganic Chemistry and Molecular Engineering, College of Chemistry and Molecular Engineering, Peking University , Beijing 100871, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Chen
authors.affiliation: Beijing National Laboratory for Molecular Sciences, MOE Key Laboratory of Bioorganic Chemistry and Molecular Engineering, College of Chemistry and Molecular Engineering, Peking University , Beijing 100871, China.
authors.forename: Meiping
authors.initials: M
authors.lastname: Zhao
mesh_terms: D015342:DNA Probes / Q000737:chemistry*; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000032:analysis* / Q000097:blood / Q000378:metabolism; D057075:Enzyme Assays / Q000295:instrumentation* / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D012015:Reference Standards; D013997:Time Factors
chemical_list: D015342:DNA Probes; D005456:Fluorescent Dyes; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase