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Characteristics of MgATP2--dependent electrogenic proton transport in tonoplast vesicles of the facultative crassulacean-acid-metabolism plant Mesembryanthemum crystallinum L.

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Abstract

Membrane vesicles were isolated from mesophyll cells of Mesembryanthemum crystallinum in the C3 state and in the crassulacean acid metabolism (CAM) state. The distribution of ATP-hydrolysis and H+-transport activities, and the activities of hydroxypyruvate reductase and Antimycin-insensitive cytochrome-c-reductase on continuous sucrose gradients was studied. For isolations carried out routinely a discontinuous sucrose gradient (24%/37%/50%) was used. Nitrate-sensitive ATP-hydrolysis and H+-transport activities increased several-fold during the transition from C3 photosynthesis to CAM. Nitrate-sensitive ATPase showed a substrate preference for ATP with an apparent Km (MgATP2-) of 0.19–0.37 mM. In both C3 and CAM states the ATPase showed a concentration-dependent stimulation by the anions chloride and malate. However, the pH optima of the two states were different: the ATPase of C3- M. crystallinum had an optimum of pH 7.4 and that of CAM-M. crystallinum an optimum of pH 8.4. The optical probe oxonol-VI was used to demonstrate the formation of MgATP2--dependent electric-potential gradients in tonoplast vesicles.

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Abbreviations

Bistris-Pronane:

1,3-bis [tris(hydroxymethyl)-methylaminol propane

CAM:

Crassulacean acid metabolism

DIDS:

4,4-dilsothiocyano-2,2-stilbene disulfonic acid:

DTT:

dithiothreitol

ER:

endoplasmic reticulum

Hepes:

4-(2-hydroxyethyl)-1-piperazineethane sulfonic acid

HPR:

hydroxypyruvate reductase

IDPase:

inosine 5′-diphosphatase

OX-VI:

oxonol VI

Tris:

2-amino-2-(hydroxymethyl)-1,3-propanediol

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  1. Institut fur Botanik, Technische Hochschule Darmstadt, Schnittspahnstrasse 3-5, D-6100, Darmstadt, Federal Republic of Germany

    I. Struve & U. Lüttge

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  1. I. Struve
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Struve, I., Lüttge, U. Characteristics of MgATP2--dependent electrogenic proton transport in tonoplast vesicles of the facultative crassulacean-acid-metabolism plant Mesembryanthemum crystallinum L.. Planta 170, 111–120 (1987). https://doi.org/10.1007/BF00392387

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