Na+-stimulated ATPase of Alkaliphilic Halotolerant Cyanobacterium Aphanothece Halophytica Translocates Na+ into Proteoliposomes Via Na+ Uniport Mechanism

Overview

Journal BMC Biochem

Publisher Biomed Central

Specialty Biochemistry

Date 2010 Aug 10

PMID 20691102

Citations 5

Authors

Kanteera Soontharapirakkul

Aran Incharoensakdi

Affiliations

Soon will be listed here.

Abstract

Background: When cells are exposed to high salinity conditions, they develop a mechanism to extrude excess Na+ from cells to maintain the cytoplasmic Na+ concentration. Until now, the ATPase involved in Na+ transport in cyanobacteria has not been characterized. Here, the characterization of ATPase and its role in Na+ transport of alkaliphilic halotolerant Aphanothece halophytica were investigated to understand the survival mechanism of A. halophytica under high salinity conditions.

Results: The purified enzyme catalyzed the hydrolysis of ATP in the presence of Na+ but not K+, Li+ and Ca2+. The apparent Km values for Na+ and ATP were 2.0 and 1.2 mM, respectively. The enzyme is likely the F1F0-ATPase based on the usual subunit pattern and the protection against N,N'-dicyclohexylcarbodiimide inhibition of ATPase activity by Na+ in a pH-dependent manner. Proteoliposomes reconstituted with the purified enzyme could take up Na+ upon the addition of ATP. The apparent Km values for this uptake were 3.3 and 0.5 mM for Na+ and ATP, respectively. The mechanism of Na+ transport mediated by Na+-stimulated ATPase in A. halophytica was revealed. Using acridine orange as a probe, alkalization of the lumen of proteoliposomes reconstituted with Na+-stimulated ATPase was observed upon the addition of ATP with Na+ but not with K+, Li+ and Ca2+. The Na+- and ATP-dependent alkalization of the proteoliposome lumen was stimulated by carbonyl cyanide m - chlorophenylhydrazone (CCCP) but was inhibited by a permeant anion nitrate. The proteoliposomes showed both ATPase activity and ATP-dependent Na+ uptake activity. The uptake of Na+ was enhanced by CCCP and nitrate. On the other hand, both CCCP and nitrate were shown to dissipate the preformed electric potential generated by Na+-stimulated ATPase of the proteoliposomes.

Conclusion: The data demonstrate that Na+-stimulated ATPase from A. halophytica, a likely member of F-type ATPase, functions as an electrogenic Na+ pump which transports only Na+ upon hydrolysis of ATP. A secondary event, Na+- and ATP-dependent H+ efflux from proteoliposomes, is driven by the electric potential generated by Na+-stimulated ATPase.

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References

Ueno S, Kaieda N, Koyama N . Characterization of a P-type Na+-ATPase of a facultatively anaerobic alkaliphile, Exiguobacterium aurantiacum. J Biol Chem. 2000; 275(19):14537-40. DOI: 10.1074/jbc.275.19.14537. View

Boekema E, van Breemen J, Brisson A, Konings W, Lolkema J . Stator structure and subunit composition of the V(1)/V(0) Na(+)-ATPase of the thermophilic bacterium Caloramator fervidus. J Mol Biol. 2000; 296(1):311-21. DOI: 10.1006/jmbi.1999.3459. View

Brown I, Fadeyev S, Kirik I, Severina I, Skulachev V . Light-dependent delta mu Na-generation and utilization in the marine cyanobacterium Oscillatoria brevis. FEBS Lett. 1990; 270(1-2):203-6. DOI: 10.1016/0014-5793(90)81268-s. View

Lerma C, Gomez-Lojero C . Preparation of a highly active ATPase of the mesophilic cyanobacterium Spirulina maxima. Photosynth Res. 2014; 11(3):265-77. DOI: 10.1007/BF00055066. View

Glynn I, Karlish S . The sodium pump. Annu Rev Physiol. 1975; 37:13-55. DOI: 10.1146/annurev.ph.37.030175.000305. View

Blumwald E, Wolosin J, Packer L . Na+/H+ exchange in the cyanobacterium Synechococcus 6311. Biochem Biophys Res Commun. 1984; 122(1):452-9. DOI: 10.1016/0006-291x(84)90497-2. View

Wiangnon K, Raksajit W, Incharoensakdi A . Presence of a Na+-stimulated P-type ATPase in the plasma membrane of the alkaliphilic halotolerant cyanobacterium Aphanothece halophytica. FEMS Microbiol Lett. 2007; 270(1):139-45. DOI: 10.1111/j.1574-6968.2007.00667.x. View

Gimmler H . Primary sodium plasma membrane ATPases in salt-tolerant algae: facts and fictions. J Exp Bot. 2000; 51(348):1171-8. View

Kaieda N, Wakagi T, Koyama N . Presence of Na(+)-stimulated V-type ATPase in the membrane of a facultatively anaerobic and halophilic alkaliphile. FEMS Microbiol Lett. 1998; 167(1):57-61. DOI: 10.1111/j.1574-6968.1998.tb13207.x. View

10.

Balnokin Y, Popova L, Andreev I . Electrogenicity of the Na+-ATPase from the marine microalga Tetraselmis (Platymonas) viridis and associated H+ countertransport. FEBS Lett. 2000; 462(3):402-6. DOI: 10.1016/s0014-5793(99)01567-7. View

11.

Neumann S, Matthey U, Kaim G, Dimroth P . Purification and properties of the F1F0 ATPase of Ilyobacter tartaricus, a sodium ion pump. J Bacteriol. 1998; 180(13):3312-6. PMC: 107283. DOI: 10.1128/JB.180.13.3312-3316.1998. View

12.

Ferguson S, Keis S, Cook G . Biochemical and molecular characterization of a Na+-translocating F1Fo-ATPase from the thermoalkaliphilic bacterium Clostridium paradoxum. J Bacteriol. 2006; 188(14):5045-54. PMC: 1539966. DOI: 10.1128/JB.00128-06. View

13.

Balnokin Y, Popova L . The ATP-driven Na(+)-pump in the plasma membrane of the marine unicellular alga, Platymonas viridis. FEBS Lett. 1994; 343(1):61-4. DOI: 10.1016/0014-5793(94)80607-1. View

14.

Reed R, Rowell P, Stewart W . Characterization of the transport of potassium ions in the cyanobacterium Anabaena variabilis Kütz. Eur J Biochem. 1981; 116(2):323-30. DOI: 10.1111/j.1432-1033.1981.tb05337.x. View

15.

Apse M, Blumwald E . Engineering salt tolerance in plants. Curr Opin Biotechnol. 2002; 13(2):146-50. DOI: 10.1016/s0958-1669(02)00298-7. View

16.

Wang H, Postier B, Burnap R . Polymerase chain reaction-based mutageneses identify key transporters belonging to multigene families involved in Na+ and pH homeostasis of Synechocystis sp. PCC 6803. Mol Microbiol. 2002; 44(6):1493-506. DOI: 10.1046/j.1365-2958.2002.02983.x. View

17.

Linnett P, Beechey R . Inhibitors of the ATP synthethase system. Methods Enzymol. 1979; 55:472-518. DOI: 10.1016/0076-6879(79)55061-7. View

18.

Allakhverdiev S, Sakamoto A, Nishiyama Y, Inaba M, Murata N . Ionic and osmotic effects of NaCl-induced inactivation of photosystems I and II in Synechococcus sp. Plant Physiol. 2000; 123(3):1047-56. PMC: 59068. DOI: 10.1104/pp.123.3.1047. View

19.

Koyama N, Koshiya K, Nosoh Y . Purification and properties of ATPase from an alkalophilic Bacillus. Arch Biochem Biophys. 1980; 199(1):103-9. DOI: 10.1016/0003-9861(80)90261-1. View

20.

Laubinger W, Dimroth P . Characterization of the Na+-stimulated ATPase of Propionigenium modestum as an enzyme of the F1F0 type. Eur J Biochem. 1987; 168(2):475-80. DOI: 10.1111/j.1432-1033.1987.tb13441.x. View