A high‐conductance mode of a poly‐3‐hydroxybutyrate/calcium/polyphosphate channel isolated from competent Escherichia coli cells

@article{Pavlov2005AHM,
  title={A high‐conductance mode of a poly‐3‐hydroxybutyrate/calcium/polyphosphate channel isolated from competent Escherichia coli cells},
  author={Evgeny V. Pavlov and Chelsey Grimbly and Catherine T M Diao and Robert J. French},
  journal={FEBS Letters},
  year={2005},
  volume={579}
}
Reconstitution into planar lipid bilayers of a poly‐3‐hydroxybutyrate/calcium/polyphosphate (PHB/Ca2+/polyP) complex from Escherichia coli membranes yields cationic‐selective, 100 pS channels (Das, S., Lengweiler, U.D., Seebach, D. and Reusch, R.N. (1997) Proof for a non‐proteinaceous calcium‐selective channel in Escherichia coli by total synthesis from (R)‐3‐hydroxybutanoic acid and inorganic polyphosphate. Proc. Natl. Acad. Sci. USA 94, 9075–9079). Here, we report that this complex can also… Expand
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The kinetic characterization of the channel in mode 2, a mode characterized by two well-defined conductance levels, a fully open state, and a major subconductance state, is reported, with asymmetric voltage-dependence indicating an asymmetry in the channel structure. Expand
Poly-3-hydroxybutyrate/polyphosphate complexes form voltage-activated Ca2+ channels in the plasma membranes of Escherichia coli.
TLDR
Bilayer patch-clamp techniques are used to demonstrate that voltage-activated Ca2+ channels composed of PHB and CaPPi are in the plasma membranes of E. coli, the first report of a biological non-proteinaceous calcium channel. Expand
Proof for a nonproteinaceous calcium-selective channel in Escherichia coli by total synthesis from (R)-3-hydroxybutanoic acid and inorganic polyphosphate.
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It is demonstrated unambiguously that the completely synthetic complex forms channels that are indistinguishable in their voltage-dependent conductance, in their selectivity for divalent cations, and in their blocking behavior from channels isolated from Escherichia coli. Expand
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TLDR
It is proposed that the polyhydroxybutyrate forms an exolipophilic-endopolarophilic helix around an inner framework helix of calcium polyphosphate that forms a channel through the membrane and may play a role in the transport of calcium, phosphate, and DNA. Expand
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Genetic competence in Escherichia coli requires poly-beta-hydroxybutyrate/calcium polyphosphate membrane complexes and certain divalent cations
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The results indicate that PHB/Ca2+/PPi complexes and the above-mentioned divalent cations each have essential but disparate roles in genetic competence. Expand
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TLDR
A protocol for transforming log-phase E. coli using dimethyl sulfoxide solutions of poly-(R)-3-hydroxybutyrate (PHB) to facilitate the transfer of plasmid DNA into cells, and certain reagents and temperature shocks to promote DNA uptake is described. Expand
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