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. Author manuscript; available in PMC: 2023 Jul 10.
Published in final edited form as: Biochemistry. 2023 Feb 6;62(4):912–922. doi: 10.1021/acs.biochem.2c00639

FIGURE 3.

FIGURE 3.

Initial velocities (ΔAbs360nm min−1) determined using a modified Invitrogen EnzChek™ assay were plotted as a function of enzyme activity without and with avH-NOX as the Fe(II)-unligated (Fe(II)-avH-NOX) or Fe(II)-NO (NO-avH-NOX) complex. The initial velocity of the cyclase activity of the AAL variant (250 nM) with and without 2.5 μM Fe(II)-avH-NOX or NO-avH-NOX is reported relative to the initial velocity of the cyclase activity of the AAL variant in the absence of avH- NOX (white bars). The initial velocity of the phosphodiesterase activity of the GGAAF variant (50 nM) with and without 0.50 μM Fe(II)-avH-NOX or NO-avH-NOX, is reported relative to the initial velocity of the phosphodiesterase activity of the GGAAF variant in the absence of avH- NOX (shaded bars). Errors bars represent the standard deviation from the mean of at least three independent trials. NS = not significant and * = p < 0.05 are for the comparison of AAL (white bars) or GGAAF (patterned bars) activities in the presence of NO-avH-NOX to those activities in the presence of Fe(II)-avH-NOX.