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13c The solution turned light blue immediately and gradually changed to purple. An excess of fresh Cr(EDTA) was added to the nitrosyl myoglobin solution at pH 10.0 to give essentially quantitative production of 2, by UV-vis. The reduced protein was repurified through a G-25 column, and the resulting solution was concentrated down by centrifugation before spectral measurements. To generate solutions of 2 at different pH, the buffer was replaced by dilution and centrifugation with desired buffer solution.
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0342286729
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note
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14b For the electrochemical reduction of nitrosyl myoglobin, monomethyl 4,4′-bipyridine was used as the mediator. A three-neck UV-vis cuvette was employed with a gold working electrode, an SCE reference electrode, and a Pt counter electrode. The solution was degassed with nitrogen, and both nitrosyl myoglobin and monomethyl 4,4′-bipyridine were added. As a -750 mV vs NHE potential was imposed, the spectra change was followed by UV-vis.
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30
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0342721815
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note
-
A methyl viologen dichloride (Aldrich) solution was made by dissolving 3 mg of methyl viologen in pH 10.0 buffer, and then it was added by aliquots to the solution of 2 in a gastight UV-vis cell. Resulting spectra are given in the Supporting Information.
-
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33
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0343591792
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note
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4Cl (Oxford).
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0342286726
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note
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2, an H-atom added, and the protein adduct allowed to minimize from several different starting conformations, resulting in the shown active-site structure.
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