In vitro synthesis and oligomerization of the mechanosensitive channel of large conductance, MscL, into a functional ion channel

Claire E. Price, Armagan Kocer, Stefan Kol, Jan Pieter van der Berg, Arnold J. M. Driessen*

*Bijbehorende auteur voor dit werk

OnderzoeksoutputAcademicpeer review

17 Citaten (Scopus)
178 Downloads (Pure)

Samenvatting

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. (C) 2010 Published by Elsevier B. V. on behalf of the Federation of European Biochemical Societies.

Originele taal-2English
Pagina's (van-tot)249-254
Aantal pagina's6
TijdschriftFEBS Letters
Volume585
Nummer van het tijdschrift1
DOI's
StatusPublished - 3-jan-2011

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