Poly-3-hydroxybutyrate/polyphosphate complexes form voltage-activated Ca2+ channels in the plasma membranes of Escherichia coli.
The lipidic polymer, poly-3-hydroxybutyrate (PHB), is found in the plasma membranes of Escherichia col complexed to calcium polyphosphate (CaPPi). The composition, location, and putative structure of the polymer salt complexes led Reusch and Sadoff (1988) to propose that the complexes function as Ca2+ channels. Here we use bilayer patch-clamp techniques to demonstrate that voltage-activated Ca2+ channels composed of PHB and CaPPi are in the plasma membranes of E. coli. Single channel calcium currents were observed in vesicles of plasma membranes incorporated into planar bilayers of synthetic 1-palmitoyl, 2-oleoyl phosphatidylcholine. The channels were extracted from cells and incorporated into bilayers, where they displayed many of the signal characteristics of protein Ca2+ channels: voltage-activated selective for divalent over monovalent cations, permeant to Ca2+, manner by La3+, Co2+, Cd2+, and Mg2+, in that order. The channel-active extract, purified by size exclusion chromatography, was found to contain only PHB and CaPPi. This composition was confirmed by the observation of comparable single channel currents with complexes reconstituted from synthetic CaPPi and PHB, isolated from E. coli. This is the first report of a biological non-proteinaceous calcium channel. We suggest that poly-3-hydroxybutyrate/calcium polyphosphate complexes are evolutionary antecedents of protein Ca2+ channels.
Department of Microbiology, Michigan State University, East Lansing 48824, USA.
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Reusch
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Huang
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D015220:Calcium Channels / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology*; D002413:Cations, Divalent; D002414:Cations, Monovalent; D002462:Cell Membrane / Q000502:physiology*; D004926:Escherichia coli / Q000502:physiology*; D006885:Hydroxybutyrates / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D008563:Membrane Lipids / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology; D008564:Membrane Potentials; D008958:Models, Molecular; D008968:Molecular Conformation; D018408:Patch-Clamp Techniques; D011091:Polyesters / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D011122:Polyphosphates / Q000737:chemistry* / Q000378:metabolism*; D000094122:Polyhydroxybutyrates
D015220:Calcium Channels; D002413:Cations, Divalent; D002414:Cations, Monovalent; D006885:Hydroxybutyrates; D008563:Membrane Lipids; D011091:Polyesters; D011122:Polyphosphates; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates