In vivo EPR on spin labeled colicin A reveals an oligomeric assembly of the pore-forming domain in E. coli membranes

被引：33

作者：

Dunkel, S. ^{[1
]}

Pulagam, L. P. ^{[1
]}

Steinhoff, H. -J. ^{[1
]}

Klare, J. P. ^{[1
]}

机构：

[1] Univ Osnabruck, Dept Phys, D-49076 Osnabruck, Germany

来源：

PHYSICAL CHEMISTRY CHEMICAL PHYSICS | 2015年 / 17卷 / 07期

关键词：

IA CHANNEL; TOPOLOGY; ELECTRON; TRANSLOCATION; REDUCTION; RESONANCE; PROTEINS; MUTANTS; HELICES; STATE;

D O I：

10.1039/c4cp05638h

中图分类号：

O64 [物理化学（理论化学）、化学物理学];

学科分类号：

070304 ; 081704 ;

摘要：

We report on the application of site-directed spin labeling (SDSL) and electron paramagnetic resonance (EPR) We report on the application of site-directed spin labeling (SDSL) and electron paramagnetic resonance (EPR) spectroscopy to study possible oligomerization of the bacterial toxin colicin A (ColA) upon membrane insertion in vitro and in vivo. We applied SDSL-EPR protocols and optimized experimental conditions to perform continuous wave EPR experiments and double electron-electron resonance distance measurements on intact Escherichia coli cells interacting with nitroxide spin-labeled ColA. Our data suggest that ColA forms dimers upon membrane insertion, thus explaining previously reported pore diameters of about 1 nm, which are unlikely to be formed by a single colicin A monomer.

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页码：4875 / 4878

页数：4

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