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In vitro synthesis and oligomerization of the mechanosensitive channel of large conductance, MscL, into a functional ion channel
Author(s) -
Price Claire E.,
Kocer Armagan,
Kol Stefan,
van der Berg Jan Pieter,
Driessen Arnold J.M.
Publication year - 2011
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2010.11.057
Subject(s) - mechanosensitive channels , ion channel , biophysics , membrane , membrane protein , chemistry , in vitro , conductance , cytoplasm , microbiology and biotechnology , biochemistry , biology , receptor , mathematics , combinatorics
Elucidation of high‐resolution structures of integral membrane proteins is drastically lagging behind that of cytoplasmic proteins. In vitro synthesis and insertion of membrane proteins into synthetic membranes could circumvent bottlenecks associated with the overexpression of membrane proteins, producing sufficient membrane‐inserted, correctly folded protein for structural studies. Using the mechanosensitive channel of large conductance, MscL, as a model protein we show that in vitro synthesized MscL inserts into YidC‐containing proteoliposomes and oligomerizes to form a homopentamer. Using planar membrane bilayers, we show that MscL forms functional ion channels capable of ion transport. These data demonstrate that membrane insertion of MscL is YidC mediated, whereas subsequent oligomerization into a functional homopentamer is a spontaneous event.