The Cu2+ transfer from PcuC·Cu1+·Cu2+ to ScoI (fig. S9B) was measured at 25°C in 20 mM Mops-NaOH (pH 7.0) and 50 mM NaCl in a SX20 stopped-flow instrument. ScoI (25 μM) was mixed with 25 μM PcuC·Cu1+·Cu2+, and Cu2+ transfer was monitored by the increase in the ScoI·Cu2+–specific absorbance at 360 nm. Three independent absorbance traces were recorded and averaged.

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