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

Price, C. E., Kocer, A., Kol, S., van der Berg, J. P. & Driessen, A. J. M., 3-Jan-2011, In : FEBS Letters. 585, 1, p. 249-254 6 p.

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  • In vitro synthesis and oligomerization of the mechanosensitive

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DOI

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.

Original languageEnglish
Pages (from-to)249-254
Number of pages6
JournalFEBS Letters
Volume585
Issue number1
Publication statusPublished - 3-Jan-2011

    Keywords

  • SecY, MscL, YidC, Reconstitution, Membrane, MEMBRANE-PROTEIN INSERTION, THIN LIPID MEMBRANES, ESCHERICHIA-COLI, INNER MEMBRANE, POLYENE ANTIBIOTICS, SINGLE RESIDUE, AMPHOTERICIN-B, SUBUNIT-C, YIDC, RECONSTITUTION

ID: 5240590