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. 2020 Jan 24;48(5):2594–2603. doi: 10.1093/nar/gkaa023

Figure 2.

Figure 2.

Functional characterization of β-hairpin loop mutants of LlaBIII. (A) Results of the DNA cleavage assay showing the nucleolytic activity of LlaBIII and its β-hairpin loop mutants (LlaBIIIK385A, LlaBIIIΔLoop and LlaBIIIPolyALA) on 1085 bp DNA with head-to-head oriented sites. (B, C) Electrophoretic mobility shift assays comparing the DNA binding affinities of WT LlaBIII and LlaBIIIΔLoop for a specific 28 bp DNA. (D–G) Comparison of the ATPase activities of (D) LlaBIII, (E) LlaBIIIK385A, (F) LlaBIIIPolyALA and (G) LlaBIIIΔLoop in presence of 46 and 28 bp specific DNA (SP_DNA) and a 44 bp non-specific DNA (NSP_DNA). 50 nM LlaBIII or its mutants, 500 nM DNA and 1 mM ATP were used for the NADH coupled ATPase assay carried out at 25°C. Shown are the averages of three replicates.