Pyrophosphate-Driven Proton Transport by Microsomal Membranes of Corn Coleoptiles
- 1 September 1985
- journal article
- research article
- Published by Oxford University Press (OUP) in Plant Physiology
- Vol. 79 (1), 159-164
- https://doi.org/10.1104/pp.79.1.159
Abstract
Corn (Zea mays L. cv. Trojan T929) coleoptile membranes were fractionated on isopycnic sucrose density gradients. Two peaks of ATP-driven H+-transport activity, corresponding to the previously characterized tonoplast (1.07 grams per cubic centimeter) and Golgi (1.13 grams per cubic centimeter) fractions (Chanson and Taiz, Plant Physiol. 1985 78: 232-240) were localized. Coincident with these were two peaks of inorganic pyrophosphate (PPi)-driven H+-transport. At saturating (3 millimolar) concentrations of Mg2+:ATP, the rate of proton transport was further enhanced by the addition of 3 millimolar PPi, and the stimulation was additive, i.e. equal to the sum of the two added separately. The specific PPi analog, imidodiphosphate, antagonized PPi-driven H+-transport, but had no effect on ATP-driven transport. Moreover, PPi-dependent proton transport in both tonoplast-enriched and Golgi-enriched fractions was strongly promoted by 50 millimolar KNO3, unlike the ATP-dependent H+-pumps of the same membranes. Taken together, the results indicate that PPi-driven proton transport is mediated by specific membrane-bound H+-translocating pyrophosphatases. Both potassium and a permanent anion (NO3- > Cl-), were required for maximum activity. The PPi-driven proton pumps were totally inhibited by N,N''-dicyclohexylcarbodiimide, but were insensitive to 100 millimolar vanadate. The PPi concentration in coleoptile extracts was determined using an NADH oxidation assay system coupled to purified pyrophosphate:fructose 6-phosphate 1-phosphotransferase (EC 2.7.1.90). The total pyrophosphate content of corn coleoptiles was 20 nanomoles/gram fresh weight. Assuming a cytoplasmic location, the calculated PPi concentration is sufficient to drive proton transport at 20% of the maximum rate measured in vitro for the tonoplast-enriched fraction, and 10% of the maximum rate for the Golgi-enriched fraction.This publication has 26 references indexed in Scilit:
- Is the Cytosolic Pi Concentration a Limiting Factor for Plant Cell Respiration?Plant Physiology, 1984
- Separation of Two Types of Electrogenic H+-Pumping ATPases from Oat RootsPlant Physiology, 1983
- Characterization of a NO3−-Sensitive H+-ATPase from Corn RootsPlant Physiology, 1983
- Anion-Sensitive, H+-Pumping ATPase in Membrane Vesicles from Oat RootsPlant Physiology, 1983
- Characterization of In Vitro Proton Pumping by Microsomal Vesicles Isolated from Corn ColeoptilesPlant Physiology, 1982
- Localization of the Proton Pump of Corn Coleoptile Microsomal Membranes by Density Gradient CentrifugationPlant Physiology, 1982
- Partial purification and properties of the proton-translocating ATPase of plant plasma membranes.Journal of Biological Chemistry, 1982
- Reconstitution of highly purified proton‐translocating pyrophosphatase from Rhodospirillum rubrumFEBS Letters, 1982
- Mitochondrial pyrophosphatase is a coupling factor of respiration and pyrophosphate synthesisFEBS Letters, 1977
- Light-induced energy conversion and the inorganic pyrophosphatase reaction in chromatophores from Rhodospirillum rubrum .1966