pH regulates cation selectivity of poly-(R)-3-hydroxybutyrate/polyphosphate channels from E. coli in planar lipid bilayers.
Poly-(R)-3-hydroxybutyrate/polyphosphate (PHB/polyP) complexes, whether isolated from the plasma membranes of bacteria or prepared from the synthetic polymers, form ion channels in planar lipid bilayers that are highly selective for Ca(2+) over Na(+) at physiological pH. This preference for divalent over monovalent cations is attributed to a high density of negative charge along the polyP backbone and the higher binding energies of divalent cations. Here we modify the charge density of polyP by varying the pH, and observe the effect on cation selectivity. PHB/polyP complexes, isolated from E. coli, were incorporated into planar lipid bilayers, and unitary current-voltage relations were determined as a function of pH. When Ca(2+) was the sole permeant cation, conductance diminished steadily from 97 +/- 6 pS at pH 7.4 to 47 +/- 3 pS at pH 5.5. However, in asymmetric solutions of Ca(2+) and Na(+), there was a moderate increase in conductance from 98 +/- 4 at pH 7.4 to 129 +/- 4 pS at pH 6.5, and a substantially larger increase to 178 +/- 6 pS at pH 5.6, signifying an increase in Na(+) permeability or disorganization of channel structure. Reversal potentials point to a sharp decrease in preference for Ca(2+) over Na(+) over a relatively small decrease in pH. Ca(2+) was strongly favored over Na(+) at physiological pH, but the channels became nonselective near the pK(2) of phosphate (approximately 6.8), and displayed weak selectivity for Na(+) over Ca(2+) at acidic pH. Evidently, PHB/polyP complexes are versatile ion carriers whose selectivity may be modulated by small adjustments of the local pH. The results may be relevant to the physiological function of PHB/polyP channels in bacteria and the role of PHB and polyP in the Streptomyces lividans potassium channel.
Das
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University of Delhi South Campus, New Delhi, India.
Reusch
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RN
D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000737:chemistry / Q000378:metabolism*; D002413:Cations, Divalent; D002414:Cations, Monovalent; D004553:Electric Conductivity; D004926:Escherichia coli / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006885:Hydroxybutyrates / Q000737:chemistry / Q000378:metabolism*; D015640:Ion Channel Gating; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism*; D008564:Membrane Potentials; D018408:Patch-Clamp Techniques; D011091:Polyesters / Q000737:chemistry / Q000378:metabolism*; D012964:Sodium / Q000378:metabolism; D015222:Sodium Channels / Q000737:chemistry / Q000378:metabolism*; D012996:Solutions; D000094122:Polyhydroxybutyrates
D002118:Calcium; D015220:Calcium Channels; D002413:Cations, Divalent; D002414:Cations, Monovalent; D006885:Hydroxybutyrates; D008051:Lipid Bilayers; D011091:Polyesters; D012964:Sodium; D015222:Sodium Channels; D012996:Solutions; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates