Single-molecule tracking of perfringolysin O assembly and membrane insertion uncoupling

被引：5
|
作者：
Senior, Michael J. T. [1 ]
Monico, Carina [1 ,2 ]
Weatherill, Eve E. [1 ,2 ]
Gilbert, Robert J. [3 ]
Heuck, Alejandro P. [4 ,5 ]
Wallace, Mark, I [2 ]
机构：
[1] Univ Oxford, Dept Chem, Chem Res Lab, Oxford, England
[2] Kings Coll London, Dept Chem, London SE1 1DB, England
[3] Univ Oxford, Wellcome Ctr Human Genet, Div Struct Biol, Oxford, England
[4] Univ Massachusetts, Dept Biochem, Amherst, MA 01003 USA
[5] Univ Massachusetts, Dept Mol Biol, Amherst, MA 01003 USA
来源：
FEBS JOURNAL | 2023年 / 290卷 / 02期
基金：
英国工程与自然科学研究理事会;
关键词：
assembly; DIB; fluorescence; PFO; photobleaching; single molecule; CHOLESTEROL-DEPENDENT CYTOLYSIN; PORE FORMATION; MECHANISM; PROTEIN; DAMAGE; FLUORESCENCE; PNEUMOLYSIN; TRANSITION; PLATFORM; BILAYERS;
D O I：
10.1111/febs.16596
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：
We exploit single-molecule tracking and optical single channel recording in droplet interface bilayers to resolve the assembly pathway and pore formation of the archetypical cholesterol-dependent cytolysin nanopore, Perfringolysin O. We follow the stoichiometry and diffusion of Perfringolysin O complexes during assembly with 60 ms temporal resolution and 20 nm spatial precision. Our results suggest individual nascent complexes can insert into the lipid membrane where they continue active assembly. Overall, these data support a model of stepwise irreversible assembly dominated by monomer addition, but with infrequent assembly from larger partial complexes.
引用
收藏
页码：428 / 441
页数：14
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