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. 2018 Aug 16;92(17):e00692-18. doi: 10.1128/JVI.00692-18

FIG 6.

FIG 6

The disulfide bond can affect the ssDNA binding abilities of PEDV nsp9. (A) The ssDNA binding abilities of nsp9-WT, nsp9-C59A, nsp9-G95E/G99E/G102E, and nsp9-C59A/G95E/G99E/G102E as determined by EMSA. Each protein was assayed at different concentrations (0, 0.41, 0.82, 1.64, 3.28, 6.56, and 10 μM) with 5 μM 5′-Cy5-labeled 36-mer ssDNA. (B) Zone-interference gel electrophoresis illustrating the association of nsp9-WT, nsp9-C59A, and nsp9- C59A/G95E/G99E/G102E with 36-mer ssDNA. The red dotted lines represent the position of the bands of nsp9-WT, nsp9-C59A, and nsp9-C59A/G95E/G99E/G102E without reaction with ssDNA. (C) The ssDNA binding affinities of nsp9-WT, nsp9-C59A, nsp9-G95E/G99E/G102E, and nsp9-C59A/G95E/G99E/G102E were measured by MST assays. Each protein was assayed in 2-fold concentration steps with 10 nM 5′-Cy5-labeled 36-mer ssDNA. The measured Kd of each protein is shown in the table.