Abstract
Continuing our work on fluorogenic substrates labeled with single fluorophores for nucleic acid modifying enzymes, here we describe the development of such substrates for DNA ligases and some base excision repair enzymes. These substrates are hairpin-type synthetic DNA molecules with a single fluorophore located on a base close to the 3' ends, an arrangement that results in strong fluorescence quenching. When such substrates are subjected to an enzymatic reaction, the position of the dyes relative to that end of the molecules is altered, resulting in significant fluorescence intensity changes. The ligase substrates described here were 5' phosphorylated and either blunt-ended or carrying short, self-complementary single-stranded 5' extensions. The ligation reactions resulted in the covalent joining of the ends of the molecules, decreasing the quenching effect of the terminal bases on the dyes. To generate fluorogenic substrates for the base excision repair enzymes formamido-pyrimidine-DNA glycosylase (FPG), human 8-oxo-G DNA glycosylase/AP lyase (hOGG1), endonuclease IV (EndoIV), and apurinic/apyrimidinic endonuclease (APE1), we introduced abasic sites or a modified nucleotide, 8-oxo-dG, at such positions that their enzymatic ex...Continue Reading
References
Dec 15, 1975·European Journal of Biochemistry·A J RaaeK Kleppe
Jul 11, 1991·Nucleic Acids Research·J Goodchild, J K Vishwanatha
May 11, 1989·Nucleic Acids Research·V Bailly, W G Verly
Jun 19, 2001·Nucleic Acids Research·E L KreklauL C Erickson
Apr 25, 2002·Nucleic Acids Research·Irina NazarenkoAyoub Rashtchian
Sep 8, 2004·Analytical Biochemistry·Sylvie SauvaigoAlain Favier
Sep 29, 2004·Current Opinion in Chemical Biology·Weihong TanTimothy J Drake
Oct 3, 2006·Angewandte Chemie·Ulrich RöschFrank Würthner
Jan 11, 2007·Biochemistry·Nikita A KuznetsovOlga S Fedorova
Nov 26, 2009·Analytical Biochemistry·Sang-Eun KimChow H Lee
Feb 10, 2011·Analytical Biochemistry·Theo T Nikiforov
Mar 15, 2011·Journal of Biomolecular Screening·Adam B ShapiroOlga Rivin
Nov 13, 2012·Talanta·Van-Trong NguyenRu-Qin Yu
May 22, 2013·Analytical Biochemistry·Graeme J ThomsonDonald J Ogilvie
Feb 5, 2014·Analytica Chimica Acta·Guillaume GinesDidier Gasparutto
Mar 19, 2014·Biosensors & Bioelectronics·Guillaume GinesDidier Gasparutto
Mar 19, 2014·Analytica Chimica Acta·Chen LiHaizhen Zhu
Jun 8, 2014·Analytical Biochemistry·Theo T Nikiforov