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Protein assemblies ejected directly from native membranes yield complexes for mass spectrometry

Protein assemblies ejected directly from native membranes yield complexes for mass spectrometry
Protein assemblies ejected directly from native membranes yield complexes for mass spectrometry
Membrane proteins reside in lipid bilayers and are typically extracted from this environment for study, which often compromises their integrity. Here we eject intact assemblies from membranes, without chemical disruption, and use mass spectrometry to define their composition. From E. coli outer membranes, we identify a chaperone-porin association and lipid interactions in the beta-barrel assembly machinery. Bridging inner and outer membranes we observe efflux pumps, and from inner membranes a pentameric pore of TonB, and the protein-conducting channel Sec YEG, in association with F1FO ATP-synthase. Intact mitochondrial membranes from Bos taurus yield respiratory complexes and fatty acid-bound dimers of the ADP/ATP transporter (ANT-1). These results highlight the importance of native membrane environments for retaining small-molecule binding, subunit interactions and associated chaperones of the membrane proteome.
0036-8075
829-834
Chorev, Dror S.
c4fd6545-5e3d-4829-ab01-138e42aaf606
Baker, Lindsay A.
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Wu, Di
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Beilsten-Edmands, Victoria
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Rouse, Sarah L.
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Zeev-Ben-Mordehai, Tzviya
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Jiko, Chimari
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Samsudin, Firdaus
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Gerle, Christoph
615fcc40-af48-4ce4-b62a-cbbad3d4c16a
Khalid, Syma
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Stewart, Alastair
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Matthews, Stephen J.
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Grunewald, Kay
a72b5170-968b-466a-a38d-b5a7e7298b4f
Robinson, Carol V.
54050df3-1bf7-42d9-a5eb-c39e010f1bbe
Chorev, Dror S.
c4fd6545-5e3d-4829-ab01-138e42aaf606
Baker, Lindsay A.
440b281a-a3c5-43a4-96e5-9d4d8a3b6add
Wu, Di
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Beilsten-Edmands, Victoria
e9cac42b-254d-473e-b4c2-1540d88d56af
Rouse, Sarah L.
67f40d9d-a7ba-49ee-a6a3-2e325ff3ad10
Zeev-Ben-Mordehai, Tzviya
47a6903a-d2dc-4014-b56c-91b1744bb40f
Jiko, Chimari
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Samsudin, Firdaus
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Gerle, Christoph
615fcc40-af48-4ce4-b62a-cbbad3d4c16a
Khalid, Syma
90fbd954-7248-4f47-9525-4d6af9636394
Stewart, Alastair
ac9a1fff-5863-4b47-8f28-a090130a7a09
Matthews, Stephen J.
ca0ec2f9-69b0-4266-b53a-937941e64560
Grunewald, Kay
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Robinson, Carol V.
54050df3-1bf7-42d9-a5eb-c39e010f1bbe

Chorev, Dror S., Baker, Lindsay A., Wu, Di, Beilsten-Edmands, Victoria, Rouse, Sarah L., Zeev-Ben-Mordehai, Tzviya, Jiko, Chimari, Samsudin, Firdaus, Gerle, Christoph, Khalid, Syma, Stewart, Alastair, Matthews, Stephen J., Grunewald, Kay and Robinson, Carol V. (2018) Protein assemblies ejected directly from native membranes yield complexes for mass spectrometry. Science, 362 (6416), 829-834. (doi:10.1126/science.aau0976).

Record type: Article

Abstract

Membrane proteins reside in lipid bilayers and are typically extracted from this environment for study, which often compromises their integrity. Here we eject intact assemblies from membranes, without chemical disruption, and use mass spectrometry to define their composition. From E. coli outer membranes, we identify a chaperone-porin association and lipid interactions in the beta-barrel assembly machinery. Bridging inner and outer membranes we observe efflux pumps, and from inner membranes a pentameric pore of TonB, and the protein-conducting channel Sec YEG, in association with F1FO ATP-synthase. Intact mitochondrial membranes from Bos taurus yield respiratory complexes and fatty acid-bound dimers of the ADP/ATP transporter (ANT-1). These results highlight the importance of native membrane environments for retaining small-molecule binding, subunit interactions and associated chaperones of the membrane proteome.

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aau0976cvrdsc_cvr27August_complete2018-fina-dsc-3 - Accepted Manuscript
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More information

Accepted/In Press date: 8 October 2018
e-pub ahead of print date: 16 November 2018
Published date: 16 November 2018

Identifiers

Local EPrints ID: 426434
URI: http://eprints.soton.ac.uk/id/eprint/426434
ISSN: 0036-8075
PURE UUID: 65f63478-ddb6-4fea-9654-3ca8a907bd7f
ORCID for Firdaus Samsudin: ORCID iD orcid.org/0000-0003-2766-4459
ORCID for Syma Khalid: ORCID iD orcid.org/0000-0002-3694-5044

Catalogue record

Date deposited: 27 Nov 2018 17:30
Last modified: 26 Nov 2021 03:05

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Contributors

Author: Dror S. Chorev
Author: Lindsay A. Baker
Author: Di Wu
Author: Victoria Beilsten-Edmands
Author: Sarah L. Rouse
Author: Tzviya Zeev-Ben-Mordehai
Author: Chimari Jiko
Author: Firdaus Samsudin ORCID iD
Author: Christoph Gerle
Author: Syma Khalid ORCID iD
Author: Alastair Stewart
Author: Stephen J. Matthews
Author: Kay Grunewald
Author: Carol V. Robinson

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