Mechanistic studies on the intramolecular one-electron transfer between the two flavins in the human neuronal nitric-oxide synthase and inducible nitric-oxide synthase flavin domains.

The Journal of Biological Chemistry
Zhi-Wen GuanTakashi Iyanagi

Abstract

Neuronal nitric-oxide synthase (nNOS) differs from inducible NOS (iNOS) in both its dependence on the intracellular Ca2+ concentration and the production rate of NO. To investigate what difference(s) exist between the two NOS flavin domains at the electron transfer level, we isolated the recombinant human NOS flavin domains, which were co-expressed with human calmodulin (CaM). The flavin semiquinones, FADH* and FMNH*, in both NOSs participate in the regulation of one-electron transfer within the flavin domain. Each semiquinone can be identified by a characteristic absorption peak at 520 nm (Guan, Z.-W., and Iyanagi, T. (2003) Arch. Biochem. Biophys. 412, 65-76). NADPH reduction of the FAD and FMN redox centers by the CaM-bound flavin domains was studied by stopped-flow and rapid scan spectrometry. Reduction of the air-stable semiquinone (FAD-FMNH*) of both domains with NADPH showed that the extent of conversion of FADH2/FMNH* to FADH*/FMNH2 in the iNOS flavin domain was greater than that of the nNOS flavin domain. The reduction of both oxidized domains (FAD-FMN) with NADPH resulted in the initial formation of a small amount of disemiquinone, which then decayed. The rate of intramolecular electron transfer between the two flavin...Continue Reading

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Citations

Aug 11, 2010·Journal of the American Chemical Society·Andrei V AstashkinChangjian Feng
Mar 23, 2010·Nitric Oxide : Biology and Chemistry·Simon Daff
Jun 16, 2004·Archives of Biochemistry and Biophysics·Jie FuTakashi Iyanagi
Aug 30, 2005·Biochemical and Biophysical Research Communications·Takashi Iyanagi
Dec 11, 2014·The FEBS Journal·Nicole G H LeferinkNigel S Scrutton
Oct 8, 2013·Nitric Oxide : Biology and Chemistry·Jing LeiTimothy R Billiar
Feb 18, 2009·Chembiochem : a European Journal of Chemical Biology·Edward BeaumontAnny Slama-Schwok
Sep 10, 2005·The Journal of Biological Chemistry·Mauro TisoDennis J Stuehr
May 27, 2005·American Journal of Physiology. Cell Physiology·Davin JagnandanDavid Fulton
Jan 12, 2021·International Journal of Biological Macromolecules·Manish ShandilyaSuman Kundu
Aug 10, 2006·Chemical Reviews·Perry A FreyGeorge H Reed
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Aug 28, 2021·International Journal of Molecular Sciences·Narimantas ČėnasLidija Kosychova

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