15N Tracing Studies on In Vitro Reactions of Ferredoxin-Dependent Nitrite Reductase and Glutamate Synthase Using Reconstituted Electron Donation Systems

Plant & Cell Physiology
Tadakatsu YoneyamaA Suzuki

Abstract

It is known that plants contain ferredoxin (Fd)-dependent nitrite reductase (NiR) and glutamate synthase (GOGAT). The Fd-NiR reaction produces ammonia from nitrite, and the activity is usually measured by nitrite disappearance. The Fd-GOGAT reaction forms two glutamates of different origin, from glutamine and 2-oxoglutarate, and the activity is measured by the oxidation of reductant (NADPH) or by formation of total glutamate. Here, a quantitative probe of the products and efficiency of the process was conducted using (15)N tracing techniques on these reactions in vitro. We quantified the reduction of (15)N-labeled [Formula: see text] to [Formula: see text] and the formation of [(15)N]glutamate and [(14)N]glutamate from [5-(15)N-amide]glutamine plus 2-oxoglutarate by NiR and GOGAT, respectively, with the reductant-Fd-NADP(+) oxidoreductase (FNR)-Fd system as the sequential electron donors. The supply of dithionite or NADPH to recombinant cyanobacterial NiR led to electron donation system-dependent formation of [(15)N]ammonium from [(15)N]nitrite. Addition of 20 mM NaCl and 20 mM Na-ascorbate accelerated nitrite reduction under high concentrations of NADPH. A sufficient supply of NADPH to recombinant Zea mays Fd-GOGAT generated c...Continue Reading

References

Apr 10, 1992·Biochimica Et Biophysica Acta·M HirasawaD B Knaff
Dec 3, 1991·Biochemistry·M A VanoniB Curti
Jan 1, 1988·Trends in Biochemical Sciences·D I Arnon
Oct 18, 1974·Nature·P J Lea, B J Miflin
Nov 1, 1974·Proceedings of the National Academy of Sciences of the United States of America·P P TrottaA Meister
Apr 1, 1970·The Biochemical Journal·D W TempestC M Brown
May 18, 1994·Biochimica Et Biophysica Acta·M HirasawaD B Knaff
Jan 1, 1995·Annual Review of Nutrition·N P Curthoys, M Watford
Jun 1, 1996·Archives of Biochemistry and Biophysics·M HirasawaD B Knaff
Feb 1, 1997·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·A K ArakakiN Carrillo
Jun 24, 2003·Journal of Molecular Biology·Robert H H van den HeuvelAndrea Mattevi
Apr 1, 1991·Plant Physiology·D HeinekeH W Heldt
Dec 29, 2007·Journal of Experimental Botany·J O GummadovaC G Bowsher

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Citations

Dec 3, 2015·Plant & Cell Physiology·Laure GaufichonAkira Suzuki

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