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^*

^*Corresponding author for this work

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
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 - 2 Feb 1998
Externally published	Yes

Bibliographical 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).

Keywords

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

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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.",

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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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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).

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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.

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

Abstract

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