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. Author manuscript; available in PMC: 2009 Aug 25.
Published in final edited form as: Biochemistry. 1998 May 19;37(20):7351–7362. doi: 10.1021/bi972864b

Figure 3.

Figure 3

Anaerobic reductive titration of N338A ETF-QO. The solution contained 20 μM R. sphaeroides N338A ETF-QO, 1.5 μM human ETF, and 0.5 μM human MCAD in 10 mM Hepes(K+) at pH 7.4. The oxidized sample (spectrum 1) was titrated with octanyl-CoA. After each addition spectra were recorded every 5-10 min until less than 0.001 au change per minute in absorbance was observed. Spectra 2 and 3 correspond to mole ratios of 0.5:1 and 1:1, respectively, and spectrum 4 is the dithionite-reduced enzyme. The inset shows the Δε430nm vs mole ratio of titrant for (●) wild-type, (+) N338T, and (◇) N338A ETF-QOs.