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

被引:32
作者
Pavlov, E [1 ]
Grimbly, C [1 ]
Diao, CTM [1 ]
French, RJ [1 ]
机构
[1] Univ Calgary, Fac Med, Dept Physiol & Biophys, Hotchkiss Brain Inst, Calgary, AB T2N 4N1, Canada
基金
加拿大健康研究院;
关键词
polyphosphate; reconstitution; planar lipid bilayers; Escherichia coli;
D O I
10.1016/j.febslet.2005.08.032
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 mu M, 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 (vertical bar V vertical bar >= 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/Ca2+/polyP complex in the membrane. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
引用
收藏
页码:5187 / 5192
页数:6
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