Fig. 4From: Mutations in the nucleotide binding and hydrolysis domains of Helicobacter pylori MutS2 lead to altered biochemical activities and inactivation of its in vivo functionNucleotide and mutation induced conformational changes in HpMutS2. a Limited proteolysis. Proteins (4 μM) either free or pre-incubated with nucleotides (1 mM) were incubated with chymotrypsin (1.25 ng) for 30 min at 37 °C. All proteolysis reactions were performed in 1X buffer A lacking magnesium. Reactions were stopped by adding a protease inhibitor cocktail and the products were separated on SDS-PAGE (10 %). Lane M: Molecular weight marker. b Thermal stability profiles of wild-type HpMutS2, HpMutS2-G338R, and HpMutS2-E413A. Proteins (1 μM) dialyzed in 50 mM Tris-Cl and 100 mM NaCl were subjected to thermal denaturation and spectra were recorded at 222 nm using Jasco-815 spectropolarimeterBack to article page