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PLANT PHYSIOLOGY , Vol 111, Issue 4 1145-1152, Copyright © 1996 by American Society of Plant Biologists
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WHOLE PLANT, ENVIRONMENTAL, AND STRESS PHYSIOLOGY |
Characterization of Glutathione Uptake in Broad Bean Leaf Protoplasts
A. Jamai, R. Tommasini, E. Martinoia and S. Delrot
Laboratoire de Physiologie et Biochimie Vegetales, Unite Associee Centre National de la Recherche Scientifique 574, Universite de Poitiers, Batiment Botanique, 40, Avenue du Recteur Pineau, 86022 Poitiers Cedex, France (A.J., S.D.)
Transport of reduced glutathione (GSH) and oxidized glutathione (GSSG) was
studied with broad bean (Vicia faba L.) leaf tissues and protoplasts.
Protoplasts and leaf discs took up GSSG at a rate about twice the uptake
rate of GSH. Detailed studies with protoplasts indicated that GSH and GSSG
uptake exhibited the same sensitivity to the external pH and to various
chemical reagents. GSH uptake was inhibited by GSSG and glutathione
conjugates. GSSG uptake was inhibited by GSH and GS conjugates, and the
uptake of metolachlor-GS was inhibited by GSSG. Various amino acids
(L-glutamic acid, L-glutamine, L-cysteine, L-glycine, L-methionine) and
peptides (glycine-glycine, glycine-glycine-glycine) affected neither the
transport of GSH nor GSSG. Uptake kinetics indicate that GSH is taken up by
a single saturable transporter, with an apparent Km of 0.4 mM, whereas GSSG
uptake exhibits two saturable phases, with an apparent Km of 7 [mu]M and
3.7 mM. It is concluded that the plasma membrane of leaf cells contains a
specific transport system for glutathione, which takes up GSSG and GS
conjugates preferentially over GSH. Proton flux measurements and
electrophysiological measurements indicate that GSH and GSSG are taken up
with proton symport. However, a detailed analysis of these measurements
suggests that the ion movements induced by GSSG differ from those induced
by GSH.
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