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. 2022 Apr 5;61(8):665–677. doi: 10.1021/acs.biochem.2c00080

Figure 7.

Figure 7

Pre-steady-state reactivity of the fully reduced ascorbate-free PHM determined by measuring the rate of appearance of the 390 nm Cu(II)–azido adduct at 390 nm using stopped-flow spectrophotometry. 500 μM of WT PHM was shot against 5 mM Ac-YVG + 20 mM sodium azide. Data were collected for 500 ms, with readings taken every 5 ms. The top trace shows the generation of the 390 nm absorbance as a function of time in seconds. The bottom trace shows the rate curve fitted to a single exponential with kobs= 72 s–1. The red dashed line represents the absorbance expected for complete reaction where all copper is oxidized to Cu(II) determined from a separate experiment where fully oxidized Cu(II) PHM was shot against buffer containing sodium azide under identical conditions.