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. Author manuscript; available in PMC: 2022 Jan 1.
Published in final edited form as: J Inorg Biochem. 2020 Nov 4;214:111298. doi: 10.1016/j.jinorgbio.2020.111298

Figure 2.

Figure 2.

Human Hsp90α protein increases superoxide (O2•−) production by nNOSµ (left panel) and nNOSα (right panel) under l-Arg depleted conditions. nNOSα protein produces larger amount of O2•− than nNOSµ (top panels), while the nNOSα-Hsp90 system generates relatively comparable BMPO-•OOH as the nNOSµ-Hsp90 (bottom panels). Specifically, Hsp90α protein enhances O2•− production by the nNOSµ and nNOSα protein at varying proportions. The EPR spin trapping measurements were performed in phosphate buffer (50 mM, pH 7.4, 0.1 mM EDTA, 0.1 mM DTPA) containing 20 nM nNOS, 3 µM of Hsp90α, 1 µM CaM, 20 mM BMPO and 0.5 mM CaCl2. The reaction was initiated by adding NADPH. The depicted EPR signals were collected at 15 minutes from the beginning of the reaction. The concentrations of nNOSα and nNOSµ were the same (20 nM) in these experiments.