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.