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A Kdp-like, high-affinity, K+-translocating ATPase is expressed during growth of Rhodobacter sphaeroides in low potassium media

Distribution of this K+-ATPase among purple non-sulphur phototrophic bacteria

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Abstract

Cells of the purple non-sulphur bacterium Rhodobacter sphaeroides express a high-affinity K+ uptake system when grown in media with low K+ concentrations. Antibodies againts the catalytic KdpB protein or the whole KdpABC complex of Escherichia coli crossreact with a 70.0 kDa R. sphaeroides protein that was expressed only in cells grown in media with low K+ concentrations. In membranes derived from R. sphaeroides cells grown with low K+ concentrations (induced cells), a high ATPase activity could be detected when assayed in Tris-HCl pH 8.0 containing 1 mM MgSO4. This ATPase activity increased upon addition of 1 mM KCl from 166 to 289 μmol ATP hydrolysed x min-1 x g protein-1 (1.7-fold stimulation). The K+-stimulated ATPase activity was inhibited approximately 93% by 0.5 mM vanadate but hardly by N,N′-dicyclohexylcarbo-diimide (DCCD). These results indicate that the inducible K+-ATPase in R. sphaeroides resembles the Kdp K+-translocating ATPase of Escherichia coli. This Kdp-like transport system is also expressed in R. capsulatus and Rhodospirillum rubrum during growth in media with low K+ concentrations suggesting a wide distribution of this transport system among phototrophic bacteria.

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Abbreviations

ΔΨ:

electrical potential difference across the cytoplasmic membrane

ΔpH:

pH difference across the cytoplasmic membrane

BSA:

bovine serum albumine

PAGE:

polyacrylamide gel electrophoresis

HEPES:

4-(2-hydroxyethyl)-1-piperazine-ethanesulfonic acid

PMSF:

phenyl-methyl-sulfonyl fluoride

DCCD:

N,N′-dicyclohexylcarbodiimide

AIB:

2-α-aminoisobutyric acid

TMG:

methyl-β-d-thiogalactopyranoside

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Author notes

  1. Tjakko Abee

    Present address: Department of Food Science, Agricultural University, Bomenweg 2, 6703 HD, Wageningen, The Netherlands

  2. Annette Siebers

    Present address: Biotechnology Laboratory, University of British Columbia, V6T 1Z3, British Columbia, Vancouver, Canada

Authors and Affiliations

  1. Department of Microbiology, University of Groningen, Kerklaan 30, 9751 NN, Haren, The Netherlands

    Tjakko Abee, Jan Knol & Wil N. Konings

  2. Department of Microbiology, University of Amsterdam, 1018 WS, Amsterdam, The Netherlands

    Klaas J. Hellingwerf

  3. Faclibereich Biologie/Chemie, Arbeitsgruppe Mikrobiologie, Universität Osnabrück, W-4500, Osnabrück, Germany

    Evert P. Bakker & Annette Siebers

Authors
  1. Tjakko Abee
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  2. Jan Knol
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  3. Klaas J. Hellingwerf
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  4. Evert P. Bakker
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  5. Annette Siebers
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  6. Wil N. Konings
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Abee, T., Knol, J., Hellingwerf, K.J. et al. A Kdp-like, high-affinity, K+-translocating ATPase is expressed during growth of Rhodobacter sphaeroides in low potassium media. Arch. Microbiol. 158, 374–380 (1992). https://doi.org/10.1007/BF00245368

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  • DOI: https://doi.org/10.1007/BF00245368

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