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Cytochemical localization of plasma membrane ATPase activity in plant cells
Authors:E Chauhan  D S Cowan  J L Hall
Institution:(1) Department of Biology, The University of Southampton, S09 3TU Southampton, UK;(2) Present address: Plant Cytochemistry and Ultrastructure Laboratory, Department of Botany, University of Delhi, Delhi, India
Abstract:Summary The cytochemical localization of ATPase activity has been investigated in maize root cells using both lead and cerium-based capture methods. With both methods, staining at the plasma membrane was observed in all cells of the root, although the precipitate obtained with cerium was more uniform and granular than that with lead. Controls using no substrate or no magnesium, beta-glycerophosphate to replace ATP, vanadate or boiled tissue generally showed little or no staining. However, biochemical studies on purified plasma membrane fractions showed that ATPase activity was markedly inhibited by fixation, particularly by glutaraldehyde, and also by lead and cerium ions. Non-enzymic hydrolysis of ATP by cerium was greater than that by lead. The value and limitations of these procedures for the localization of plasma membrane H+-ATPase activity are summarized in relation to previous criticisms of these methods.Abbreviations DTT dithiothreitol - EDTA ethylene diaminetetraacetic acid - GP B-glycerophosphate - PCMBS p-chloromercuribenzene sulphonic acid - PMSF phenylmethylsulphonyl fluoride
Keywords:ATPase  Cerium  Cytochemistry  Fixation  Zea mays roots  Plasma membrane
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