Biochemistry and Genetics of Bacterial H+-Translocating ATPases

Fillingame, Robert H.

doi:10.1016/b978-0-12-152511-8.50009-8

Cited by 136 publications

(72 citation statements)

References 191 publications

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“…The membrane-bound ATPase of M. barkeri described here was similar to' common FoF, ATPases in the following properties: (i) divalent cation requirement, (ii) DCCD' inhibition in the membrane-bound form, (iii) solubilization of the ATPase from the membranes by incubation in a low-ionic-strength buffer with EDTA, and (iv) rebinding of the solubilized ATPase'to depleted membrane in a high-ionic-strength buffer with Mg2+. It is apparent that the acid-induced ATP synthesis observed in whole cells of M. barkeri (23) ATPase consists of five different subunits with a stoichiometry of 3a:3P:-y:8:e (9). On the other hand, ATPases with a more simple subunit structure have also been reported; the F1 from Bacillus megaterium (19) and Bacillus subtilis (22) had only two subunits, and Clostridium pasteurianum (4) had four subunits as an FoF1 complex.…”

Section: Resultsmentioning

confidence: 99%

Characterization and purification of the membrane-bound ATPase of the archaebacterium Methanosarcina barkeri

Inatomi¹

1986

J Bacteriol

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Membrane-bound ATPase was found in membranes of the archaebacterium Methanosarcina barkeri. The ATPase activity required divalent cations, Mg2' or Mn2 , and maximum activity was obtained at pH 5.2. The activity was specifically stimulated by HS03-with a shift of optimal pH to 5.8, and N,N'-dicyclohexylcarbodiimide inhibited ATP hydrolysis. The enzyme could be solubilized from membranes by incubation in 1 mM Tris-maleate buffer (pH 6.9) containing 0.5 mM EDTA. The solubilized ATPase was purified by DEAE-Sepharose and Sephacryl S-300 chromatography. The molecular weight of the purified enzyme was estimated to be 420,000 by gel filtration through Sephacryl S-300. Polyacrylamide gel electrophoresis in sodium dodecyl sulfate revealed two classes of subunit, Mr 62,000 (a) and 49,000 (Ii) associated in the molar ratio 1:1. These results suggest that the ATPase of M. barkeri is similar to the FoF, type ATPase found in many eubacteria.Methanogenic bacteria are strictly anaerobic organisms, and most species are able to grow autotrophically with H2 and CO2 as substrates for methanogenesis. On the basis of studies of the 16S RNA sequence, they are considered to belong to the archaebacteria (1). Other properties of methanogens are also different from those of the eubacteria; the membranes of methanogens contain ether-linked polyisoprenoid glycerol lipids (30) and their cell walls lack peptidoglycan (14). Apart from methanogens, the archaebacteria include extreme halophiles and thermoacidophiles.In the chemiosmotic mechanism of Mitchell (20), a proton motive force established across the cell membrane is a driving force for ATP synthesis. An H+-translocating ATPase (FoF1 ATPase) which catalyzes ATP synthesis has been observed in membranes of both eucaryotes and eubacteria (26). Although membrane-bound ATPase activities were found in membranes of thermoacidophilic archaebacteria, Thermoplasma (28), and Sulfolobus (32) Solubilization and purification of the ATPase. The membrane fraction was suspended in 1 mM Tris-maleate buffer (pH 6.9) containing 0.5 mM EDTA. After incubation at 8°C for 1 h with gentle stirring, the suspension was centrifuged at 100,000 x g for 90 min. The supernatant (42 ml, 2.4 mg of protein per ml) was applied to a DEAE-Sepharose column (2.6 by 20 cm) equilibrated with 50 mM Tris-maleate (pH 6.9) (column buffer). After the column had been washed with one column volume of the column buffer, a linear gradient from 0 to 0.6 M NaCl in 50 mM Tris-maleate buffer (pH 6.9) was run at 8°C. The fractions containing ATPase activity were pooled (15 ml), and solid ammonium sulfate was added up to reach 65% saturation. The precipitate obtained after centrifugation at 10,000 x g for 20 min was dissolved in a minimal volume of 50 mM Tris-maleate buffer (pH 6.9), and the solution was centrifuged to remove any insoluble material. The supernatant was then applied to a Sephacryl S-300 column (1.6 by 80 cm) equilibrated with the column buffer. The flow rate was 10 ml/min at 8°C, and the fractions with ATPase activity were collec...

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Section: Resultsmentioning

confidence: 99%

Characterization and purification of the membrane-bound ATPase of the archaebacterium Methanosarcina barkeri

Inatomi¹

1986

J Bacteriol

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“…The FIF 0 ATPases of bovine mitochondria and Escherichia coli have many common structural feal tures (reviews [1][2][3][4][5]). They appear to be made up of a membrane-bound domain Fl which is attached to a transmembrane structure, F0.…”

Section: Introductionmentioning

confidence: 99%

Subunit equivalence in Escherichia coli and bovine heart mitochondrial F₁F₀ ATPases

1982

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“…In addition, available gene (unc or arp) sequence data have revealed significant homology. Several reviews on the structure of bacterial H+-ATPase have recently been published (2,28,89,90,103), and procedures for purification of the enzyme have been presented (3, 89). Detailed genetic studies have also been reviewed (28,29,104).…”

Section: Proton-translocating Atpasementioning

confidence: 99%

pH Homeostasis in Lactic Acid Bacteria

Hutkins¹,

Nannen

1993

Journal of Dairy Science

244

157

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The ability of lactic acid bacteria to regulate their cytoplasmic or intracellular pH is one of the most important physiological requirements of the cells. Cells unable to maintain a near neutral intracellular pH during growth or storage at low extracellular pH may lose viability and cellular activity. Despite the importance of pH homeostasis in the lactic acid bacteria, however, an understanding of cytoplasmic pH regulation has only recently begun to emerge. This review describes the specific effects of low pH on lactic acid bacteria, reports recent research on the physiological role of intracellular pH as a regulator of various metabolic activities in lactic acid bacteria, and presents the means by which lactic acid bacteria defend against low intracellular pH. Particular attention is devoted to the proton-translocating ATPase, an enzyme that is largely responsible for pH homeostasis in fermentative lactic acid bacteria. (Key words: pH, pH homeostasis, lactic acid bacteria) Abbreviation key: pHout = extracellular pH, pHi, = intracellular pH, ApH = pH gradient, H+-ATPase = proton-translocating ATPase.

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Biochemistry and Genetics of Bacterial H+-Translocating ATPases

Cited by 136 publications

References 191 publications

Characterization and purification of the membrane-bound ATPase of the archaebacterium Methanosarcina barkeri

Characterization and purification of the membrane-bound ATPase of the archaebacterium Methanosarcina barkeri

Subunit equivalence in Escherichia coli and bovine heart mitochondrial F₁F₀ ATPases

pH Homeostasis in Lactic Acid Bacteria

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Biochemistry and Genetics of Bacterial H+-Translocating ATPases

Cited by 136 publications

References 191 publications

Characterization and purification of the membrane-bound ATPase of the archaebacterium Methanosarcina barkeri

Characterization and purification of the membrane-bound ATPase of the archaebacterium Methanosarcina barkeri

Subunit equivalence in Escherichia coli and bovine heart mitochondrial F1F0 ATPases

pH Homeostasis in Lactic Acid Bacteria

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Subunit equivalence in Escherichia coli and bovine heart mitochondrial F₁F₀ ATPases