An in vivo screen reveals protein-lipid interactions crucial for gating a mechanosensitive channel

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The bacterial mechanosensitive channel MscL is the best-studied mechanosensor, thus serving as a paradigm of how a protein senses and responds to mechanical force. Models for the transition of Escherichia coli MscL from closed to open states propose a tilting of the transmembrane domains in the plane of the membrane, suggesting dynamic protein-lipid interactions. Here, we used a rapid in vivo assay to assess the function of channels that were post-translationally modified at several different sites in a region just distal to the cytoplasmic end of the second transmembrane helix. We utilized multiple probes with various affinities for the membrane environment. The in vivo functional data, combined with site-directed mutagenesis, single-channel analyses, and tryptophan fluorescence measurements, confirmed that lipid interactions within this region are critical for MscL gating. The data suggest a model in which this region acts as an anchor for the transmembrane domain tilting during gating. Furthermore, the conservation of analogous motifs among many other channels suggests a conserved protein-lipid dynamic mechanism.

Original languageEnglish (US)
Pages (from-to)694-702
Number of pages9
JournalFASEB Journal
Volume25
Issue number2
DOIs
StatePublished - Feb 2011

Fingerprint

Lipids
Membranes
Mutagenesis
Proteins
Site-Directed Mutagenesis
Anchors
Tryptophan
Escherichia coli
Assays
Conservation
Fluorescence

Keywords

  • Methanethiosulfonate modification
  • MscL
  • Osmoregulation

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Genetics
  • Molecular Biology

Cite this

An in vivo screen reveals protein-lipid interactions crucial for gating a mechanosensitive channel. / Iscla, Irene; Wray, Robin; Blount, Paul.

In: FASEB Journal, Vol. 25, No. 2, 02.2011, p. 694-702.

Research output: Contribution to journalArticle

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