Fig. 4 | Nature Communications

Fig. 4

From: Evolution of a General RNA-Cleaving FANA Enzyme

Fig. 4

Functional analysis of FANAzymes. a Structure–activity profile of NGS12-1 showing functional mutations (green) and non-functional mutations (red) observed in highly abundant NGS sequences. b Monovalent and bivalent metal ion requirements for NGS12-7. c Mn2+ or Ca2+ activity profile of NGS12-7. d Pre-steady state kinetic plots of NGS12-7 in the presence of 2 mM MnCl2 or 10 mM CaCl2 in buffer (pH 8.5) containing 200 mM NaCl at 24 °C ([Fz] = 2.5 µM, [S] = 0.5 µM). e, f Substrate specificity of NGS12-7. e Engineered versions of NGS12-7 in which the substrate-binding arms were modified to recognize different RNA substrates. f Gel images showing the cleavage activity of engineered NGS12-7 variants toward different RNA substrates. Reactions were performed in buffer (pH 8.5) containing 200 mM NaCl supplemented with the labeled bivalent metal ion at 24 °C (Fz) = 2.5 µM, [S] = 0.5 µM). S substrate, P product

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