TY - JOUR
T1 - A high-conductance mode of a poly-3-hydroxybutyrate/calcium/polyphosphate channel isolated from competent Escherichia coli cells
AU - Pavlov, Evgeny
AU - Grimbly, Chelsey
AU - Diao, Catherine T.M.
AU - French, Robert J.
N1 - Funding Information:
This work was supported by an operating grant to R.J.F. from the Canadian Institutes of Health Research (CIHR). We thank Dr. Rosetta Reusch for critical comments, advice, and encouragement, and Dr. Eleonora Zakharian for assistance with preliminary experiments.
PY - 2005/9/26
Y1 - 2005/9/26
N2 - 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 form larger, weakly selective pores, with a maximal conductance ranging from 250 pS to 1 nS in different experiments (symmetric 150 mM KCl). Single channels were inhibited by lanthanum (IC50 = 42 ± 4 μM, means ± S.E.M.) with an unusually high Hill coefficient (8.4 ± 1.2). Transition to low-conductance states (<250 pS) was favored by increased membrane polarization (|V| ≥ 50 mV). High conductance states (>250 pS) may reflect conformations important for genetic transformability, or "competence", of the bacterial cells, which requires the presence of the PHB/Ca 2+/polyP complex in the membrane.
AB - 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 form larger, weakly selective pores, with a maximal conductance ranging from 250 pS to 1 nS in different experiments (symmetric 150 mM KCl). Single channels were inhibited by lanthanum (IC50 = 42 ± 4 μM, means ± S.E.M.) with an unusually high Hill coefficient (8.4 ± 1.2). Transition to low-conductance states (<250 pS) was favored by increased membrane polarization (|V| ≥ 50 mV). High conductance states (>250 pS) may reflect conformations important for genetic transformability, or "competence", of the bacterial cells, which requires the presence of the PHB/Ca 2+/polyP complex in the membrane.
KW - Escherichia coli
KW - Planar lipid bilayers
KW - Polyphosphate
KW - Reconstitution
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U2 - 10.1016/j.febslet.2005.08.032
DO - 10.1016/j.febslet.2005.08.032
M3 - Article
C2 - 16150446
AN - SCOPUS:24944588768
SN - 0014-5793
VL - 579
SP - 5187
EP - 5192
JO - FEBS Letters
JF - FEBS Letters
IS - 23
ER -