Allosteric activation of an ion channel triggered by modification of mechanosensitive nano-pockets

Charalampos Kapsalis; Bolin Wang; Hassane El Mkami; Samantha Jane Pitt; Jason Schnell; Terry K. Smith; Jonathan Lippiat; Bela E. Bode; Christos Pliotas

doi:10.1038/s41467-019-12591-x

Allosteric activation of an ion channel triggered by modification of mechanosensitive nano-pockets

Charalampos Kapsalis, Bolin Wang, Hassane El Mkami, Samantha Jane Pitt, Jason Schnell, Terry K. Smith, Jonathan Lippiat, Bela E. Bode, Christos Pliotas

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

Abstract

Lipid availability within transmembrane nano-pockets of ion channels is linked with mechanosensation. However, the effect of hindering lipid-chain penetration into nano-pockets on channel structure has not been demonstrated. Here we identify nano-pockets on the large conductance mechanosensitive channel MscL, the high-pressure threshold channel. We restrict lipid-chain access to the nano-pockets by mutagenesis and sulfhydryl modification, and monitor channel conformation by PELDOR/DEER. For a single site located at the entrance of the nano-pockets and distal to the channel pore we generate an allosteric response in the absence of tension. Single-channel recordings reveal a decrease in the pressure activation threshold of the modified channel and a sub-conducting state in the absence of applied tension. Threshold is restored to wild-type levels upon reduction of the sulfhydryl modification. The modification associated with the conformational change restricts lipid access to the nano-pocket, and interrupts the contact between the membrane and the channel that mediates mechanosensitivity.

Original language	English
Article number	4619
Number of pages	14
Journal	Nature Communications
Volume	10
DOIs	https://doi.org/10.1038/s41467-019-12591-x
Publication status	Published - 10 Oct 2019

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Understanding sensitivity gains in pulse: Understanding sensitivity gains in pulse EPR on multimeric membrane proteins
Bode, B. E. (PI) & Pliotas, C. (CoI)
The Leverhulme Trust
14/05/19 → 18/02/22
Project: Standard
Cryogen-free arbitrary waveform EPR: Cryogen-free arbitrary waveform EPR for structural Biology and Biophysics
Bode, B. E. (PI), Lovett, J. E. (CoI), Pliotas, C. (CoI), Schwarz-Linek, U. (CoI), Smith, G. M. (CoI), Stewart, A. J. (CoI) & White, M. (CoI)
1/05/18 → 30/04/19
Project: Standard
State of the art pulse EPR instrumentati: State of the art pulse EPR instrumentation for long range distance measurements in biomacromolecules
Smith, G. M. (PI), Bode, B. E. (CoI), Naismith, J. (CoI), Schiemann, O. (CoI) & White, M. (CoI)
The Wellcome Trust
1/09/12 → 31/08/17
Project: Standard

Bela Ernest Bode
- beb2st-andrews.acuk
Person: Academic
Samantha J. Pitt
- sjp24st-andrews.acuk
Person: Academic

Data Underpinning - Allosteric activation of an ion channel triggered by modification of mechanosensitive nano-pockets
Kapsalis, C. (Creator), Wang, B. (Creator), El Mkami, H. (Creator), Pitt, S. J. (Creator), Schnell, J. (Creator), Smith, T. K. (Creator), Lippiat, J. (Creator), Bode, B. E. (Creator) & Pliotas, C. (Creator), University of St Andrews, 2019
DOI: 10.17630/fa8518b0-13ba-4b11-bad4-e5c6b000738d
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Cite this

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title = "Allosteric activation of an ion channel triggered by modification of mechanosensitive nano-pockets",

abstract = "Lipid availability within transmembrane nano-pockets of ion channels is linked with mechanosensation. However, the effect of hindering lipid-chain penetration into nano-pockets on channel structure has not been demonstrated. Here we identify nano-pockets on the large conductance mechanosensitive channel MscL, the high-pressure threshold channel. We restrict lipid-chain access to the nano-pockets by mutagenesis and sulfhydryl modification, and monitor channel conformation by PELDOR/DEER. For a single site located at the entrance of the nano-pockets and distal to the channel pore we generate an allosteric response in the absence of tension. Single-channel recordings reveal a decrease in the pressure activation threshold of the modified channel and a sub-conducting state in the absence of applied tension. Threshold is restored to wild-type levels upon reduction of the sulfhydryl modification. The modification associated with the conformational change restricts lipid access to the nano-pocket, and interrupts the contact between the membrane and the channel that mediates mechanosensitivity.",

author = "Charalampos Kapsalis and Bolin Wang and {El Mkami}, Hassane and Pitt, {Samantha Jane} and Jason Schnell and Smith, {Terry K.} and Jonathan Lippiat and Bode, {Bela E.} and Christos Pliotas",

note = "C.P. is supported by the Royal Society of Edinburgh. C.P. acknowledges support by Tenovus (T15/41) and Carnegie Trust (OS000256) and the University of St Andrews, the University of Leeds and the Chinese Scholarship Council for the C.K. and B.W. studentships respectively. S.J.P. is supported by the Royal Society of Edinburgh. J.S. acknowledges funding from the MRC (M019152 and L018578). B.E.B. and C.P. acknowledge support by the Leverhulme Trust (RPG-2018-397). This work was supported by Wellcome Trust [099149/Z/12/Z] and BBSRC equipment grants (BB/R013780/1).",

year = "2019",

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day = "10",

doi = "10.1038/s41467-019-12591-x",

language = "English",

volume = "10",

journal = "Nature Communications",

issn = "2041-1723",

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T1 - Allosteric activation of an ion channel triggered by modification of mechanosensitive nano-pockets

AU - Kapsalis, Charalampos

AU - Wang, Bolin

AU - El Mkami, Hassane

AU - Pitt, Samantha Jane

AU - Schnell, Jason

AU - Smith, Terry K.

AU - Lippiat, Jonathan

AU - Bode, Bela E.

AU - Pliotas, Christos

N1 - C.P. is supported by the Royal Society of Edinburgh. C.P. acknowledges support by Tenovus (T15/41) and Carnegie Trust (OS000256) and the University of St Andrews, the University of Leeds and the Chinese Scholarship Council for the C.K. and B.W. studentships respectively. S.J.P. is supported by the Royal Society of Edinburgh. J.S. acknowledges funding from the MRC (M019152 and L018578). B.E.B. and C.P. acknowledge support by the Leverhulme Trust (RPG-2018-397). This work was supported by Wellcome Trust [099149/Z/12/Z] and BBSRC equipment grants (BB/R013780/1).

PY - 2019/10/10

Y1 - 2019/10/10

N2 - Lipid availability within transmembrane nano-pockets of ion channels is linked with mechanosensation. However, the effect of hindering lipid-chain penetration into nano-pockets on channel structure has not been demonstrated. Here we identify nano-pockets on the large conductance mechanosensitive channel MscL, the high-pressure threshold channel. We restrict lipid-chain access to the nano-pockets by mutagenesis and sulfhydryl modification, and monitor channel conformation by PELDOR/DEER. For a single site located at the entrance of the nano-pockets and distal to the channel pore we generate an allosteric response in the absence of tension. Single-channel recordings reveal a decrease in the pressure activation threshold of the modified channel and a sub-conducting state in the absence of applied tension. Threshold is restored to wild-type levels upon reduction of the sulfhydryl modification. The modification associated with the conformational change restricts lipid access to the nano-pocket, and interrupts the contact between the membrane and the channel that mediates mechanosensitivity.

AB - Lipid availability within transmembrane nano-pockets of ion channels is linked with mechanosensation. However, the effect of hindering lipid-chain penetration into nano-pockets on channel structure has not been demonstrated. Here we identify nano-pockets on the large conductance mechanosensitive channel MscL, the high-pressure threshold channel. We restrict lipid-chain access to the nano-pockets by mutagenesis and sulfhydryl modification, and monitor channel conformation by PELDOR/DEER. For a single site located at the entrance of the nano-pockets and distal to the channel pore we generate an allosteric response in the absence of tension. Single-channel recordings reveal a decrease in the pressure activation threshold of the modified channel and a sub-conducting state in the absence of applied tension. Threshold is restored to wild-type levels upon reduction of the sulfhydryl modification. The modification associated with the conformational change restricts lipid access to the nano-pocket, and interrupts the contact between the membrane and the channel that mediates mechanosensitivity.

U2 - 10.1038/s41467-019-12591-x

DO - 10.1038/s41467-019-12591-x

M3 - Article

SN - 2041-1723

VL - 10

JO - Nature Communications

JF - Nature Communications

M1 - 4619

ER -

Allosteric activation of an ion channel triggered by modification of mechanosensitive nano-pockets

Abstract

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Projects

Understanding sensitivity gains in pulse: Understanding sensitivity gains in pulse EPR on multimeric membrane proteins

Cryogen-free arbitrary waveform EPR: Cryogen-free arbitrary waveform EPR for structural Biology and Biophysics

State of the art pulse EPR instrumentati: State of the art pulse EPR instrumentation for long range distance measurements in biomacromolecules

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Bela Ernest Bode

Samantha J. Pitt

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Data Underpinning - Allosteric activation of an ion channel triggered by modification of mechanosensitive nano-pockets

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