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. 1987 Dec;85(4):996–999. doi: 10.1104/pp.85.4.996

Active Glucose Transport and Proton Pumping in Tonoplast Membrane of Zea mays L. Coleoptiles Are Inhibited by Anti-H+-ATPase Antibodies

Thomas Rausch 1,2,3,1, Dennis N Butcher 1,2,3, Lincoln Taiz 1,2,3
PMCID: PMC1054382  PMID: 16665844

Abstract

A tonoplast enriched fraction was obtained from Zea mays L. coleoptiles by isopycnic centrifugation of microsomal membranes in a sucrose step gradient. At the 18/26% interface chloride-stimulated and nitrate-inhibited proton pumping activity coincided with a Mg2+-ATP dependent accumulation of 3-O-methyl-d-glucose (OMG) as determined by a membrane filtration technique using 14C-labeled substrate. OMG transport showed an apparently saturable component with a Km of 110 micromolar, and was completely inhibited by 10 micromolar carbonyl cyanide m-chlorophenylhydrazone. Polyclonal antibodies against solubilized native tonoplast H+-ATPase and its 62 and 72 kilodalton subunits were assayed for their ability to inhibit proton pumping and OMG accumulation. Antibodies against both the native enzyme and the putative catalytic subunit (72 kilodalton) strongly inhibited proton pumping and OMG transport whereas antibodies against the 62 kilodalton subunit had only a slight effect on both processes.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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