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. Author manuscript; available in PMC: 2016 Jul 1.
Published in final edited form as: Mol Microbiol. 2015 May 15;97(2):263–280. doi: 10.1111/mmi.13022

FIGURE 3. S18-1 protein binds Zn2+.

FIGURE 3

A. SDS-PAGE of purified proteins. Lanes: 1) S18-1 dialyzed in presence of Zn2+, 2) S18-1 dialyzed in presence of EDTA, 3) S18-2 dialyzed in presence of Zn2+, 4) S18-2 dialyzed in presence of EDTA; arrows depict 10 kDa and 15 kDa marker position

B. Zinc ion release: Protein preparations mixed with the zinc-binding dye MF-2 were exposed to hydrogen peroxide and fluorescence at Ex/Em (325 nm/490 nm) was measured over time.

C. S18-1-Zn2+ binding curve: MF-2 dye was used in a competition assay with S18-1 protein to measure the fraction of Zn2+ bound to S18-1 and calculate the dissociation constant (Kd). Error bars are ±1 standard deviation for technical replicates for both axes.