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

Alvin CK 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

7 Citations (Scopus)

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
Pages (from-to)1-10
Number of pages10
JournalScientific Reports
Volume9
Early online date12 Feb 2019
DOIs
Publication statusE-pub ahead of print - 12 Feb 2019

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

Cite this

Teo, Alvin CK ; 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. pp. 1-10.
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Teo, ACK, Lee, SC, Pollock, NL, Stroud, Z, Hall, S, Thakker, A, Pitt, AR, Dafforn, TR, Spickett, CM & Roper, DI 2019, 'Analysis of SMALP co-extracted phospholipids shows distinct membrane environments for three classes of bacterial membrane protein', Scientific Reports, vol. 9, 1813, pp. 1-10. https://doi.org/10.1038/s41598-018-37962-0

Analysis of SMALP co-extracted phospholipids shows distinct membrane environments for three classes of bacterial membrane protein. / Teo, Alvin CK; 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, 1813, 12.02.2019, p. 1-10.

Research output: Contribution to journalArticle

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

AU - Teo, Alvin CK

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

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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.

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