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Figure 2
(a) Thermal stability of ENDO proteins determined by differential scanning fluorimetry (DSF). The melting temperatures (Tm) of ENDO-WT, ENDO-D118A and ENDO-D88A (75 µM; P) with or without the indicated divalent cation (at 0.5 mM; P+Mg, P+Mn, P+Mg+Mn) and the compounds DPBA (1) (P+Mg+Mn+DPBA) and L-742,001 (2) (P+Mg+Mn+L-742,001) (at 450 µM, ligand:protein ratio = 6) were measured in a thermofluorescence experiment. (b) Polyacrylamide/8 M urea gels of inhibition of endonuclease activity by DPBA (1) and L-742,001 (2). Increasing concentrations of molecules (1) and (2) were incubated with 20 µM protein and 1 µM single-stranded RNA. The reaction products were analyzed in 20% polyacrylamide/8 M urea gels. The lane labelled NC lacked protein in the reaction mixture, while the lane labelled EDTA contained all reagents plus 5 mM EDTA.

Volume 5| Part 2| March 2018| Pages 223-235
ISSN: 2052-2525