Figure 3. WAH-1 is a monomer under reduced conditions and a dimer under oxidized conditions.
(A) WAH-1 was eluted as a monomer (60 kDa) in Superdex 200 (S-200) size exclusion chromatography in reduced conditions. The chromatographic fractions from the S-200 column were analyzed by SDS-PAGE (bottom panel). (B) WAH-1 was eluted in two peaks (oxidized dimer and monomer) in the presence of 2.5 mM H2O2. (C) The nuclease activity of CPS-6 was reduced in the oxidized condition, but was enhanced by WAH-1. (D) The reduced monomeric form of WAH-1 significantly enhanced the nuclease activity of oxidized CPS-6 in plasmid DNA nicking assays. The mean percentages with standard errors shown at the bottom of the gel were calculated from three independent experiments.