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Medientyp:
E-Artikel
Titel:
An H⁺-ATPase Assay: Proton Pumping and ATPase Activity Determined Simultaneously in the Same Sample
Beteiligte:
Palmgren, Michael Gjedde
Erschienen:
American Society of Plant Physiologists, 1990
Erschienen in:
Plant Physiology, 94 (1990) 3, Seite 882-886
Sprache:
Englisch
ISSN:
0032-0889;
1532-2548
Entstehung:
Anmerkungen:
Beschreibung:
A continuous spectrophotometric assay of H+-ATPase activity was developed by combining two well-known methods for measuring proton pumping and ATPase activity. Proton uptake into plasma membrane vesicles from Avena sativa L. (cv Rhiannon) was monitored as the absorbance decrease at 495 nm of the ΔpH probe acridine orange. Simultaneously, ATPase activity was measured by following the absorbance decrease at 340 nanometers by coupling ATP hydrolysis enzymatically to the oxidation of NADH. This H+-ATPase assay is convenient for determining the relative relationship between ATP hydrolysis and proton pumping.