Endonuclease activity of FEN-1-XPG hybrid protein on flap and bubble substrates

<p><b>Copyright information:</b></p><p>Taken from "Domain swapping between FEN-1 and XPG defines regions in XPG that mediate nucleotide excision repair activity and substrate specificity"</p><p></p><p>Nucleic Acids Research 2007;35(9):3053-3063.</p><p>Published online 22 Apr 2007</p><p>PMCID:PMC1888826.</p><p>© 2007 The Author(s)</p> () Purified proteins were incubated for 45 min at 30°C with 2.5 nM of flap substrate (lanes 1–8) in the presence of 0.5 mM MnCl for XPG and 1 mM MnCl for FEN-1-XPG and FEN-1. The following protein concentrations were used: 0 nM (lane 1), 0.5 nM (lane 2), 1 nM (lane 3), 2 nM (lane 4), 4 nM (lane 5), 8 nM (lane 6), 20 nM (lane 7) and 40 nM (lane 8). The position of the 5′P label on the substrate is indicated by the asterisk. The length of the products was determined by comparison with an authentic standard and is indicated. () Incision activity of purified proteins on the bubble substrate using the conditions described in . () Cleavage efficiency of XPG, FEN-1 and FEN-1-XPG on the flap substrate (left) and bubble substrate (right) with increasing protein concentration. Graphs represent the quantification of two independent experiments. Standard deviations are represented by error bars