Figure 2.

Zn²⁺ Binding and displacement of Zn²⁺/As³⁺ from ZRANB2 zfm apo-peptides using spectrophotometric quantification of free Zn²⁺. A, Increasing amounts of each ZRANB2 apo-peptide (0-5 μM) was incubated with 10 μM Zn²⁺ for 30 minutes at room temperature. Increase in peptide bound Zn²⁺ with increase in peptide concentration shows Zn²⁺ binding to the peptides. B, 5 μM of each ZRANB2 apo-peptide was initially incubated with 10 μM Zn²⁺ for 30 minutes at room temperature, followed by titration with increasing concentration of As³⁺ (0-5 μM) for 30 minutes at room temperature. Decrease in peptide bound Zn²⁺ with increase in As³⁺ concentration shows displacement of Zn²⁺ by As³⁺ from each peptide. C, 5 μM of each ZRANB2 apo-peptide was initially incubated with 5 μM As³⁺ for 30 minutes at room temperature, followed by titration with increasing concentration of Zn²⁺ (0-10 μM) for 30 minutes at room temperature. Increase in peptide bound Zn²⁺ with increase in Zn²⁺ concentration shows displacement of As³⁺ by Zn²⁺ from each peptide. Open circles represent ZNF1 while open squares represent ZNF2 for all the panels. Each point in each curve represents the Mean ± SD from three independent experiments.