Analysis of SMALP co-extracted phospholipids shows distinct membrane environments for three classes of bacterial membrane protein

Alvin C. K. Teo, Sarah C. Lee, Naomi L. Pollock, Zoe Stroud, Stephen Hall, Alpesh Thakker, Andrew R. Pitt, Timothy R. Dafforn, Corinne M. Spickett, David I. Roper

Research output: Contribution to journalArticle

Abstract

Biological characterisation of membrane proteins lags behind that of soluble proteins. This reflects issues with the traditional use of detergents for extraction, as the surrounding lipids are generally lost, with adverse structural and functional consequences. In contrast, styrene maleic acid (SMA) copolymers offer a detergent-free method for biological membrane solubilisation to produce SMA-lipid particles (SMALPs) containing membrane proteins together with their surrounding lipid environment. We report the development of a reverse-phase LC-MS/MS method for bacterial phospholipids and the first comparison of the profiles of SMALP co-extracted phospholipids from three exemplar bacterial membrane proteins with different topographies: FtsA (associated membrane protein), ZipA (single transmembrane helix), and PgpB (integral membrane protein). The data showed that while SMA treatment per se did not preferentially extract specific phospholipids from the membrane, SMALP-extracted ZipA showed an enrichment in phosphatidylethanolamines and depletion in cardiolipins compared to the bulk membrane lipid. Comparison of the phospholipid profiles of the 3 SMALP-extracted proteins revealed distinct lipid compositions for each protein: ZipA and PgpB were similar, but in FtsA samples longer chain phosphatidylglycerols and phosphatidylethanolamines were more abundant. This method offers novel information on the phospholipid interactions of these membrane proteins.

Original languageEnglish
Article number1813
JournalScientific Reports
Volume9
Issue number1
DOIs
Publication statusPublished - 12 Feb 2019

Fingerprint

Bacterial Proteins
Phospholipids
Membrane Proteins
Lipids
Styrene
Membranes
Phosphatidylethanolamines
Membrane Lipids
Detergents
Phosphatidylglycerols
Proteins
Cardiolipins
maleic acid

Bibliographical note

© The Author(s) 2019. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction
in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if
changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If
material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain
permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

Cite this

Teo, Alvin C. K. ; Lee, Sarah C. ; Pollock, Naomi L. ; Stroud, Zoe ; Hall, Stephen ; Thakker, Alpesh ; Pitt, Andrew R. ; Dafforn, Timothy R. ; Spickett, Corinne M. ; Roper, David I. / Analysis of SMALP co-extracted phospholipids shows distinct membrane environments for three classes of bacterial membrane protein. In: Scientific Reports. 2019 ; Vol. 9, No. 1.
@article{4926d1a69af24e519de47a53e1f92188,
title = "Analysis of SMALP co-extracted phospholipids shows distinct membrane environments for three classes of bacterial membrane protein",
abstract = "Biological characterisation of membrane proteins lags behind that of soluble proteins. This reflects issues with the traditional use of detergents for extraction, as the surrounding lipids are generally lost, with adverse structural and functional consequences. In contrast, styrene maleic acid (SMA) copolymers offer a detergent-free method for biological membrane solubilisation to produce SMA-lipid particles (SMALPs) containing membrane proteins together with their surrounding lipid environment. We report the development of a reverse-phase LC-MS/MS method for bacterial phospholipids and the first comparison of the profiles of SMALP co-extracted phospholipids from three exemplar bacterial membrane proteins with different topographies: FtsA (associated membrane protein), ZipA (single transmembrane helix), and PgpB (integral membrane protein). The data showed that while SMA treatment per se did not preferentially extract specific phospholipids from the membrane, SMALP-extracted ZipA showed an enrichment in phosphatidylethanolamines and depletion in cardiolipins compared to the bulk membrane lipid. Comparison of the phospholipid profiles of the 3 SMALP-extracted proteins revealed distinct lipid compositions for each protein: ZipA and PgpB were similar, but in FtsA samples longer chain phosphatidylglycerols and phosphatidylethanolamines were more abundant. This method offers novel information on the phospholipid interactions of these membrane proteins.",
author = "Teo, {Alvin C. K.} and Lee, {Sarah C.} and Pollock, {Naomi L.} and Zoe Stroud and Stephen Hall and Alpesh Thakker and Pitt, {Andrew R.} and Dafforn, {Timothy R.} and Spickett, {Corinne M.} and Roper, {David I.}",
note = "{\circledC} The Author(s) 2019. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.",
year = "2019",
month = "2",
day = "12",
doi = "10.1038/s41598-018-37962-0",
language = "English",
volume = "9",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",
number = "1",

}

Analysis of SMALP co-extracted phospholipids shows distinct membrane environments for three classes of bacterial membrane protein. / Teo, Alvin C. K.; Lee, Sarah C.; Pollock, Naomi L.; Stroud, Zoe; Hall, Stephen; Thakker, Alpesh; Pitt, Andrew R.; Dafforn, Timothy R.; Spickett, Corinne M.; Roper, David I.

In: Scientific Reports, Vol. 9, No. 1, 1813, 12.02.2019.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Analysis of SMALP co-extracted phospholipids shows distinct membrane environments for three classes of bacterial membrane protein

AU - Teo, Alvin C. K.

AU - Lee, Sarah C.

AU - Pollock, Naomi L.

AU - Stroud, Zoe

AU - Hall, Stephen

AU - Thakker, Alpesh

AU - Pitt, Andrew R.

AU - Dafforn, Timothy R.

AU - Spickett, Corinne M.

AU - Roper, David I.

N1 - © The Author(s) 2019. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PY - 2019/2/12

Y1 - 2019/2/12

N2 - Biological characterisation of membrane proteins lags behind that of soluble proteins. This reflects issues with the traditional use of detergents for extraction, as the surrounding lipids are generally lost, with adverse structural and functional consequences. In contrast, styrene maleic acid (SMA) copolymers offer a detergent-free method for biological membrane solubilisation to produce SMA-lipid particles (SMALPs) containing membrane proteins together with their surrounding lipid environment. We report the development of a reverse-phase LC-MS/MS method for bacterial phospholipids and the first comparison of the profiles of SMALP co-extracted phospholipids from three exemplar bacterial membrane proteins with different topographies: FtsA (associated membrane protein), ZipA (single transmembrane helix), and PgpB (integral membrane protein). The data showed that while SMA treatment per se did not preferentially extract specific phospholipids from the membrane, SMALP-extracted ZipA showed an enrichment in phosphatidylethanolamines and depletion in cardiolipins compared to the bulk membrane lipid. Comparison of the phospholipid profiles of the 3 SMALP-extracted proteins revealed distinct lipid compositions for each protein: ZipA and PgpB were similar, but in FtsA samples longer chain phosphatidylglycerols and phosphatidylethanolamines were more abundant. This method offers novel information on the phospholipid interactions of these membrane proteins.

AB - Biological characterisation of membrane proteins lags behind that of soluble proteins. This reflects issues with the traditional use of detergents for extraction, as the surrounding lipids are generally lost, with adverse structural and functional consequences. In contrast, styrene maleic acid (SMA) copolymers offer a detergent-free method for biological membrane solubilisation to produce SMA-lipid particles (SMALPs) containing membrane proteins together with their surrounding lipid environment. We report the development of a reverse-phase LC-MS/MS method for bacterial phospholipids and the first comparison of the profiles of SMALP co-extracted phospholipids from three exemplar bacterial membrane proteins with different topographies: FtsA (associated membrane protein), ZipA (single transmembrane helix), and PgpB (integral membrane protein). The data showed that while SMA treatment per se did not preferentially extract specific phospholipids from the membrane, SMALP-extracted ZipA showed an enrichment in phosphatidylethanolamines and depletion in cardiolipins compared to the bulk membrane lipid. Comparison of the phospholipid profiles of the 3 SMALP-extracted proteins revealed distinct lipid compositions for each protein: ZipA and PgpB were similar, but in FtsA samples longer chain phosphatidylglycerols and phosphatidylethanolamines were more abundant. This method offers novel information on the phospholipid interactions of these membrane proteins.

UR - https://doi.org/10.17036/researchdata.aston.ac.uk.00000393

UR - http://www.nature.com/articles/s41598-018-37962-0

UR - http://www.scopus.com/inward/record.url?scp=85061504775&partnerID=8YFLogxK

U2 - 10.1038/s41598-018-37962-0

DO - 10.1038/s41598-018-37962-0

M3 - Article

VL - 9

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

IS - 1

M1 - 1813

ER -