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. Author manuscript; available in PMC: 2011 May 26.

Published in final edited form as: Biochemistry. 2007 Dec 11;47(1):92–100. doi: 10.1021/bi701859s

Anaerobic reductive titration of Y501F ETF-QO. The solution contained ~30 μM R. sphaeroides Y501F ETF-QO, 2 μM human ETF, and 2 μM MCAD in 20 mM Hepes(K⁺) at pH 7.4. The oxidized sample (spectrum 1) was titrated with octanyl-CoA. Spectra were recorded every 5–10 minutes until less than 0.001 a.u. change per minute in absorbance was observed. The inset shows the Δε_{433 nm} vs. mole ratio of titrant for (○) native, ( ) Y501F, ( ) T525A, and ( ) Y501F/T525A. Spectra 2 and 3 correspond to mole ratios of 0.5:1 and 1:1, respectively, and spectrum 4 is the dithionite reduced sample of Y501F ETF-QO.

Inline graphic — Anaerobic reductive titration of Y501F ETF-QO. The solution contained ~30 μM R. sphaeroides Y501F ETF-QO, 2 μM human ETF, and 2 μM MCAD in 20 mM Hepes(K⁺) at pH 7.4. The oxidized sample (spectrum 1) was titrated with octanyl-CoA. Spectra were recorded every 5–10 minutes until less than 0.001 a.u. change per minute in absorbance was observed. The inset shows the Δε_{433 nm} vs. mole ratio of titrant for (○) native, ( ) Y501F, ( ) T525A, and ( ) Y501F/T525A. Spectra 2 and 3 correspond to mole ratios of 0.5:1 and 1:1, respectively, and spectrum 4 is the dithionite reduced sample of Y501F ETF-QO.