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. Author manuscript; available in PMC: 2018 Jul 5.
Published in final edited form as: J Am Chem Soc. 2017 Jun 22;139(26):8828–8836. doi: 10.1021/jacs.7b01212

Figure 3.

Figure 3

Metal selectivity of the His6 site ascertained by the B-ΔHis3Asp pull-down assay. The concentrations of Ni(II) and Zn(II) in the supernatant of each sample were determined by ICP- MS. B-ΔHis3Asp (10 μM) was incubated with 10 μM Ni(II), and/or Zn(II) for 72 h at 37 °C in 75 mM HEPES, 100 mM NaCl, 2 mM CaCl2, pH 7.0 and the mixture was treated with streptavidin agarose resin. (A) Ni(II) was added first and the Ni(II) + B-ΔHis3Asp mixture was incubated for 30 min at room temperature prior to addition of Zn(II). (B) Zn(II) was added first. The mean and SDM are reported (n = 4 for samples with B-ΔHis3Asp n added, n = 2 for samples without B-ΔHis3Asp).