Transmembrane electron transport in sealed and NAD(P)H‐loaded right‐side‐out plasma membrane vesicles isolated from maize (Zea mays L.) roots

Electron transport across plasma membranes has been observed in vivo in several plant species and tissues after the application of ferricyanide (hexacyanoferrate III, HCF III). In the present work, a transmembrane electron flow was demonstrated in sealed and NAD(P)H–loaded right‐side‐out (apoplastic...

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Published inJournal of experimental botany Vol. 55; no. 401; pp. 1343 - 1349
Main Authors Menckhoff, Mathias, Lüthje, Sabine
Format Journal Article
LanguageEnglish
Published Oxford Oxford University Press 01.06.2004
Oxford Publishing Limited (England)
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Summary:Electron transport across plasma membranes has been observed in vivo in several plant species and tissues after the application of ferricyanide (hexacyanoferrate III, HCF III). In the present work, a transmembrane electron flow was demonstrated in sealed and NAD(P)H–loaded right‐side‐out (apoplastic‐side‐out) plasma membrane vesicles isolated from maize (Zea mays L.) roots. HCF III was reduced at a rate of up to 126 nmol min–1 mg–1 protein by NADPH‐loaded vesicles, while reduction rates with NADH‐loaded vesicles were several‐fold lower. Coincident with the reduction of HCF III, NAD(P)H oxidation was observed inside the vesicles. The dependence of reduction on K+ indicated an electrogenic transmembrane electron flow. Application of 100 µM calcium decreased HCF III reduction up to 66%, while pre‐incubation with 200 µM warfarin or diphenylene iodonium inhibited transmembrane electron transport only weakly. Fe3+‐EDTA was not reduced significantly by NADPH‐loaded plasma membrane vesicles, whereas XTT was reduced at a rate of 765 pmol min–1 mg–1 protein. The results suggested a major function for NADPH in transmembrane electron flow and were discussed in conjunction with in vivo experiments.
Bibliography:To whom correspondence should be addressed. Fax: +49 40 42816 254. E‐mail: s.luthje@botanik.uni‐hamburg.de
 Abbreviations: BPDS, bathophenanthroline disulphonate; Brij 58, polyoxyethylene 20 cetyl ether; DPI, diphenylene iodonium; FRE, ferric reductase; FRO, ferric reductase‐oxidase; HCF III, ferricyanide; hexacyanoferrate III; ISO, inside‐out; ROS, right‐side‐out; XTT, 2,3‐bis[2‐methoxy‐4‐nitro‐5‐sulphophenyl]‐2H‐tetrazolium‐5‐carboxanilide.
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Received 12 November 2003; Accepted 19 March 2004
local:erh155
ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:0022-0957
1460-2431
DOI:10.1093/jxb/erh155