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. Author manuscript; available in PMC: 2014 Nov 19.
Published in final edited form as: Biochemistry. 2013 Nov 8;52(46):8323–8332. doi: 10.1021/bi401305w

Figure 2.

Figure 2

Determination of the redox potential of the proximal disulfide in sfALR substituted with 5-deaza-FAD. Panel A shows the spectrum of 20 μM 5-deaza-FAD enzyme (see Experimental Procedures) in 50 mM phosphate buffer, pH 7.0, containining 1 mM EDTA recorded before and immediately after the addition of 5 mM DTT (solid and dashed lines respectively). Panel B shows the difference spectrum dominated at 439 nm by the blue shift in the leading edge of the oxidized 5-deazaflavin absorbance. Panel C follows the extent of these changes at 439 nm as a function of the redox poise of mixtures of reduced and oxidized DTT (see Experimental Procedures).