In vitro reconstitution of electron transport from glucose-6-phosphate and NADPH to nitrite

Abstract:

:An NADPH-dependent NO2--reducing system was reconstituted in vitro using ferredoxin (Fd) NADP+ oxidoreductase (FNR), Fd, and nitrite reductase (NiR) from the green alga Chlamydomonas reinhardtii. NO2- reduction was dependent on all protein components and was operated under either aerobic or anaerobic conditions. NO2- reduction by this in vitro pathway was inhibited up to 63% by 1 mm NADP+. NADP+ did not affect either methyl viologen-NiR or Fd-NiR activity, indicating that inhibition was mediated through FNR. When NADPH was replaced with a glucose-6-phosphate dehydrogenase (G6PDH)-dependent NADPH-generating system, rates of NO2- reduction reached approximately 10 times that of the NADPH-dependent system. G6PDH could be replaced by either 6-phosphogluconate dehydrogenase or isocitrate dehydrogenase, indicating that G6PDH functioned to: (a) regenerate NADPH to support NO2- reduction and (b) consume NADP+, releasing FNR from NADP+ inhibition. These results demonstrate the ability of FNR to facilitate the transfer of reducing power from NADPH to Fd in the direction opposite to that which occurs in photosynthesis. The rate of G6PDH-dependent NO2- reduction observed in vitro is capable of accounting for the observed rates of dark NO3- assimilation by C. reinhardtii.

journal_name

Plant Physiol

journal_title

Plant physiology

authors

Jin T,Huppe HC,Turpin DH

doi

10.1104/pp.117.1.303

subject

Has Abstract

pub_date

1998-05-01 00:00:00

pages

303-9

issue

1

eissn

0032-0889

issn

1532-2548

journal_volume

117

pub_type

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