Characterization of Na+- atpase in the halotolerant cyanobacterium aphanothece halophytica
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Title Characterization of Na+- atpase in the halotolerant cyanobacterium aphanothece halophytica
Creator Kanjana Wiangnon
Contributor Aran Incharoensakdi
Publisher Chulalongkorn University
Publication Year 2548
Keyword Sodium ions, Cyanobacteria, Salinity
Abstract Extrusion of sodium ion is needed to balance and maintain sodium level for microorganisms to survive in hypersalin environment. Activity of Na+- ATPase in halotolerant cyanobacterium aphanothece haolphytica was investigated. Cells were preloaded with 22Na+ and the extrusion of 22Na+ from cells was followed by energizing the cells with glucose. To ascertain that this Na+ extrusion is Na+- ATPase, the plasma membranes vesicles were prepared by aqueous polymer two-phase partitioning with dextran and polyethylene glycol and used for the assay of Na+- ATPase activity by measuring ATP hydrolysis. The ATPase activities were stimulated to greatest extent in the presence of 100 mM Na+ and 5 mM Mg2+ ions, pH 7.6. The K[subscript m] value for ATP and Na+ ions were 1.66 mM and 25 mM respectively. The maximum velocities (V [subscript max])were 0.66 and 0.50 umol/min/mg protein using ATP and Na+ as variable substrate respectively. The activity was inhibited by vanadate, a potent inhibitor of P-type ATPase. The activity was decreased about half of original value by 0.01 mM socium vanadate and 0.20 mM gramicidin. Alternatively, the membrane vesicle containing Na+- ATPase, exhibited 22Na+ transport by the addition of ATP, which was inhibited by sodium ionophore and gramicidin. It is likely that the Na+- ATPase belongs to P-type and is involved in Na+ transport. The enzyme activity was enhanced by the high salinity of growth medium.
ISBN 9745327786
URL Website cuir.car.chula.ac.th
Chulalongkorn University

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