Skip to main content
. Author manuscript; available in PMC: 2010 May 19.
Published in final edited form as: Biochemistry. 2009 May 19;48(19):4019–4030. doi: 10.1021/bi900103d

Fig. 4. Effects of conservative mutations on UvrD1 activities.

Fig. 4

(A) Aliquots (3 µg) of recombinant wild-type (WT) UvrD1 and the indicated mutants were analyzed by SDS-PAGE. The Coomassie blue-stained gels are shown. The sizes (kDa) and positions of marker proteins are indicated. (B) ATPase specific activity was determined as specified under Methods. Each datum is the average of three separate UvrD1 titration experiments; error bars denote the standard deviation. (C) Helicase reactions were performed as described under Methods. Complete reaction mixtures contained 1 mM ATP, 50 nM 32P-labeled tailed duplex DNA substrate, 75 ng Ku and 100 ng of wild-type or mutant UvrD1 protein as specified. The products were analyzed by native PAGE and visualized by autoradiography. Reactions without added protein (lane --), with wild-type UvrD1 only (–Ku) or with Ku only (–UvrD1) were included as controls. A reaction lacking protein that was heat denatured prior to PAGE is shown in lane Δ.