Helicity, membrane incorporation, orientation and thermal stability of the large conductance mechanosensitive ion channel from E. coli

Isaiah T. Arkin; Sergei I. Sukharev; Paul Blount; Ching Kung; Axel T. Brünger

doi:10.1016/S0005-2736(97)00219-8

Helicity, membrane incorporation, orientation and thermal stability of the large conductance mechanosensitive ion channel from E. coli

Isaiah T. Arkin, Sergei I. Sukharev, Paul Blount, Ching Kung, Axel T. Brünger

Research output: Contribution to journal › Article › peer-review

24 Scopus citations

Abstract

In this report, we present studies on the large conductance mechanosensitive ion channel (MscL) from E. coli in detergent micelles and lipid vesicle. Both transmission Fourier transform infrared spectroscopy and circular dichroism (CD) spectra indicate that the protein is highly helical in detergent as well as liposomes. The secondary structure of the proteins was shown to be highly resistant towards denaturation (25-95°C) based on an ellipticity thermal profile. Amide H⁺/D⁺ exchange was shown to be extensive (ca. 66%), implying that two thirds of the protein are water accessible. MscL, reconstituted in oriental lipid bilayers, was shown to possess a net bilayer orientation using dichroic ratios measured by attenuated total- reflection Fourier transform infrared spectroscopy. Here, we present and discuss this initial set of structural data on this new family of ion- channel proteins.

Original language	English (US)
Pages (from-to)	131-140
Number of pages	10
Journal	Biochimica et Biophysica Acta - Biomembranes
Volume	1369
Issue number	1
DOIs	https://doi.org/10.1016/S0005-2736(97)00219-8
State	Published - Feb 2 1998

Keywords

Circular dichroism
Fourier transform infrared spectroscopy (FT-IR)
Ion channel
Lipid bilayer
Membrane protein

ASJC Scopus subject areas

Biophysics
Biochemistry
Cell Biology

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10.1016/S0005-2736(97)00219-8

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title = "Helicity, membrane incorporation, orientation and thermal stability of the large conductance mechanosensitive ion channel from E. coli",

abstract = "In this report, we present studies on the large conductance mechanosensitive ion channel (MscL) from E. coli in detergent micelles and lipid vesicle. Both transmission Fourier transform infrared spectroscopy and circular dichroism (CD) spectra indicate that the protein is highly helical in detergent as well as liposomes. The secondary structure of the proteins was shown to be highly resistant towards denaturation (25-95°C) based on an ellipticity thermal profile. Amide H+/D+ exchange was shown to be extensive (ca. 66%), implying that two thirds of the protein are water accessible. MscL, reconstituted in oriental lipid bilayers, was shown to possess a net bilayer orientation using dichroic ratios measured by attenuated total- reflection Fourier transform infrared spectroscopy. Here, we present and discuss this initial set of structural data on this new family of ion- channel proteins.",

keywords = "Circular dichroism, Fourier transform infrared spectroscopy (FT-IR), Ion channel, Lipid bilayer, Membrane protein",

author = "Arkin, {Isaiah T.} and Sukharev, {Sergei I.} and Paul Blount and Ching Kung and Br{\"u}nger, {Axel T.}",

note = "Funding Information: This work was funded in part by a grant to Axel T. Briinger from NIH (GM-54160-01) and a grant to Sergei Sukharev from NASA (NAGW-4934).",

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T1 - Helicity, membrane incorporation, orientation and thermal stability of the large conductance mechanosensitive ion channel from E. coli

AU - Arkin, Isaiah T.

AU - Sukharev, Sergei I.

AU - Blount, Paul

AU - Kung, Ching

AU - Brünger, Axel T.

N1 - Funding Information: This work was funded in part by a grant to Axel T. Briinger from NIH (GM-54160-01) and a grant to Sergei Sukharev from NASA (NAGW-4934).

PY - 1998/2/2

Y1 - 1998/2/2

N2 - In this report, we present studies on the large conductance mechanosensitive ion channel (MscL) from E. coli in detergent micelles and lipid vesicle. Both transmission Fourier transform infrared spectroscopy and circular dichroism (CD) spectra indicate that the protein is highly helical in detergent as well as liposomes. The secondary structure of the proteins was shown to be highly resistant towards denaturation (25-95°C) based on an ellipticity thermal profile. Amide H+/D+ exchange was shown to be extensive (ca. 66%), implying that two thirds of the protein are water accessible. MscL, reconstituted in oriental lipid bilayers, was shown to possess a net bilayer orientation using dichroic ratios measured by attenuated total- reflection Fourier transform infrared spectroscopy. Here, we present and discuss this initial set of structural data on this new family of ion- channel proteins.

AB - In this report, we present studies on the large conductance mechanosensitive ion channel (MscL) from E. coli in detergent micelles and lipid vesicle. Both transmission Fourier transform infrared spectroscopy and circular dichroism (CD) spectra indicate that the protein is highly helical in detergent as well as liposomes. The secondary structure of the proteins was shown to be highly resistant towards denaturation (25-95°C) based on an ellipticity thermal profile. Amide H+/D+ exchange was shown to be extensive (ca. 66%), implying that two thirds of the protein are water accessible. MscL, reconstituted in oriental lipid bilayers, was shown to possess a net bilayer orientation using dichroic ratios measured by attenuated total- reflection Fourier transform infrared spectroscopy. Here, we present and discuss this initial set of structural data on this new family of ion- channel proteins.

KW - Circular dichroism

KW - Fourier transform infrared spectroscopy (FT-IR)

KW - Ion channel

KW - Lipid bilayer

KW - Membrane protein

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JO - Biochimica et Biophysica Acta - Biomembranes

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Helicity, membrane incorporation, orientation and thermal stability of the large conductance mechanosensitive ion channel from E. coli

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