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PLANT PHYSIOLOGY , Vol 106, Issue 2 731-737, Copyright © 1994 by American Society of Plant Biologists

METABOLISM AND ENZYMOLOGY

Direct Measurement of ATP-Dependent Proton Concentration Changes and Characterization of a K+-Stimulated ATPase in Pea Chloroplast Inner Envelope Vesicles

R. Shingles and R. E. McCarty
Department of Biology, The Johns Hopkins University, Baltimore, Maryland 21218-2685

Inner envelope membrane vesicles prepared from pea (Pisum sativum L. var Laxton's Progress No. 9) chloroplasts have K+-stimulated ATPase activity with a pH optimum of 8.4. ATP addition to inner envelope vesicles loaded with pyranine caused a decrease in pyranine fluorescence that was consistent with internal acidification. The transmembrane pH change induced by the addition of 5 mM ATP was about 0.4 unit. Measurement of phosphate released by ATP hydrolysis paralleled the pH change, indicating that intravesicular acidification was linked to ATPase activity. Vanadate, molybdate, N-ethylmaleimide, and dithiothreitol inhibited ATP-dependent vesicle acidification completely, whereas ATPase activity was only partially inhibited. These data indicate that pea chloroplast inner envelope vesicles contain a proton translocating ATPase and that the pyranine-loading method can be utilized to study directly ATP-dependent H+ transport across these membranes.


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R. Shingles, M. North, and R. E. McCarty
Ferrous Ion Transport across Chloroplast Inner Envelope Membranes
Plant Physiology, March 1, 2002; 128(3): 1022 - 1030.
[Abstract] [Full Text] [PDF]

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M. H. Roh, R. Shingles, M. J. Cleveland, and R. E. McCarty
Direct Measurement of Calcium Transport across Chloroplast Inner-Envelope Vesicles
Plant Physiology, December 1, 1998; 118(4): 1447 - 1454.
[Abstract] [Full Text]


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Copyright © 1994 by the American Society of Plant Biologists