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Planta

, Volume 174, Issue 3, pp 349–357 | Cite as

Sugar nucleotides dissipate ATP-generated transmembrane pH gradient in Golgi vesicles from suspension-cell protoplasts ofChenopodium rubrum L.

  • Maria Gogarten-Boekels
  • Johann Peter Gogarten
  • Friedrich-Wilhelm Bentrup
Article

Abstract

A microsomal vesicle fraction (GV) markedly enriched by the Golgi marker enzyme latent inosine diphosphatase (IDPase) has been isolated from photoautotrophic suspension-cell protoplasts ofChenopodium rubrum L. Addition of ATP creates a substantial pH gradient across the GV membrane as measured by accumulation of acridine orange. The GV showed a density of 1.14 g·cm-3 by equilibrium density centrifugation on sucrose gradients. Coincidence of acridine-orange accumulation and IDPase activity was confirmed on Percoll gradients. Formation of the pH gradient half-saturates at 0.3 mM MgATP, peaks at pH 7, and is competitively inhibited by ADP (ki≤0.1 mM), but not by Pi; it is hardly inhibited by orthovanadate, quickly dissipated by monensink2=18 nM), nigericin (k1/2=25 nM), and sluggishly by N-ethylmaleimide (k1/2≈35 μM). Inhibition by KNO3 (k1/2≈6.7 mM) is incomplete (60%). Uridine 5′-diphosphate (UDP)-glucose, UDP-galactose, but not UDP-mannose and the pertinent sugars, dissipate the ATP-generated pH gradient (k1/2≈10–20 mM UDP-glucose; optimum pH at 7.8). This UDP-glucose activity is accompanied by release of Pi, but not of glucose or sucrose. UDP-glucoseinduced Pi release from the GV saturates (k1/2=1 mM UDP-glucose; optimum pH at 7) and is completely inhibited by the anion-channel blocker 4,4′-diisothiocyano-2,2′-stilbene disulfonic acid (DIDS;k1/2=140 μM). The GV incorporates UDP-[U-14C]glucose into an acid-labile, alkaline-stable macromolecular compound; this process is like-wise inhibited by DIDS. We propose a model including, inter alia, a UDP-glucose/uridine-5′-monophosphate translocator and a phosphate-permeable anion channel to operate in Golgi vesicles ofChenopodium rubrum.

Key words

Chenopodium Golgi vesicle Proton ATPase Protoplast Sugar nucleotide Suspension cell 

Abbreviations

AO

acridine orange

DIDS

4,4′-diisothiocyano-2,2′-stilbene disulfonic acid

FCCP

carbonyl cyanidep-trifluoromethyoxyphenyl hydrazone

GV

Golgi-vesicle-enriched microsomal fraction

IDPase

mosine diphosphatase

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Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • Maria Gogarten-Boekels
    • 1
  • Johann Peter Gogarten
    • 1
  • Friedrich-Wilhelm Bentrup
    • 1
  1. 1.Botanisches Institut I der Justus-Liebig-UniversitätGiessenFederal Republic of Germany

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