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. Author manuscript; available in PMC: 2010 Jun 21.
Published in final edited form as: Biochemistry. 2009 May 12;48(18):3864–3876. doi: 10.1021/bi8021087

FIGURE 4.

FIGURE 4

Fluorescence intensities of the FMN subdomain in various nNOS protein constructs. (A) Fluorescence intensities of fully-oxidized FMNCaM, FMNCaMoxy, and nNOSr proteins, and free FMN were determined at the indicated concentrations by excitation at 457 nm and monitoring emission at 525 nm. The fluorescence intensities of the FNR subdomain were also obtained to subtract its contribution from the total flavin fluorescence intensity in the nNOSr. (B) Fluorescence intensity versus the concentration slope value for each protein obtained by linear regression analysis of the data in panel A. The slope of the FNR subdomain is depicted as a black bar that is superimposed onto the bar value for nNOSr. It was then subtracted to obtain an FMN-specific fluorescence value for nNOSr. Inset, scale-up plot of the FMN-specific fluorescence values for each nNOS protein. Data are representative of at least two experiments.