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Fig. 1 | BMC Microbiology

Fig. 1

From: Mutations in the nucleotide binding and hydrolysis domains of Helicobacter pylori MutS2 lead to altered biochemical activities and inactivation of its in vivo function

Fig. 1

a Schematic representation of HpMutS2 variants used in this study. Walker-A and Walker-B motifs of HpMutS2 were identified by multiple sequence alignment of MutS2 proteins from different bacteria using Clustal Omega (http://www.ebi.ac.uk/Tools/msa/clustalo/). The mutations in the walker-A and Walker-B motifs were introduced using site directed mutagenesis. The mutated amino acids are highlighted in red. All the proteins were C-terminally His6-tagged. The LDLK motif and Smr domain are conserved nuclease sites of HpMutS2. b Effect of DNA substrates on ATPase activity of HpMutS2. HpMutS2 (45 nM) was incubated with increasing concentrations of ATP [0, 10, 20, 40, 60, 80, 100, 200, 400, 600, 800, and 1000 (μM)]. DNA substrates (1 μM) were added separately to the reaction mixtures. After incubation at 37 °C for 30 min the reactions were stopped by EDTA (50 mM) and the products were separated by TLC. ATP [γ-32P] was used as tracer to monitor the product formation. Reaction velocities were calculated by quantifying the proportion of products formed to un-reacted substrate divided by incubation time

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