Using a SMALP platform to determine a sub-nm single particle cryo-EM membrane protein structure

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Using a SMALP platform to determine a sub-nm single particle cryo-EM membrane protein structure. / Parmar, Mayuriben; Rawson, Shaun; Scarff, Charlotte A.; Goldman, Adrian; Dafforn, Timothy R.; Muench, Stephen P.; Postis, Vincent L. G.

In: Biochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 1860, No. 2, 02.02.2018, p. 378-383.

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Parmar, Mayuriben ; Rawson, Shaun ; Scarff, Charlotte A. ; Goldman, Adrian ; Dafforn, Timothy R. ; Muench, Stephen P. ; Postis, Vincent L. G. / Using a SMALP platform to determine a sub-nm single particle cryo-EM membrane protein structure. In: Biochimica et Biophysica Acta (BBA) - Biomembranes. 2018 ; Vol. 1860, No. 2. pp. 378-383.

Bibtex

@article{7996c65d6acd42feb773ff93374755c7,
title = "Using a SMALP platform to determine a sub-nm single particle cryo-EM membrane protein structure",
abstract = "The field of membrane protein structural biology has been revolutionized over the last few years with a number of high profile structures being solved using cryo-EM including Piezo, Ryanodine receptor, TRPV1 and the Glutamate receptor. Further developments in the EM field hold the promise of even greater progress in terms of greater resolution, which for membrane proteins is still typically within the 4–7 {\AA} range. One advantage of a cryo-EM approach is the ability to study membrane proteins in more “native” like environments for example proteoliposomes, amphipols and nanodiscs. Recently, styrene maleic acid co-polymers (SMA) have been used to extract membrane proteins surrounded by native lipids (SMALPs) maintaining a more natural environment. We report here the structure of the Escherichia coli multidrug efflux transporter AcrB in a SMALP scaffold to sub-nm resolution, with the resulting map being consistent with high resolution crystal structures and other EM derived maps. However, both the C-terminal helix (TM12) and TM7 are poorly defined in the map. These helices are at the exterior of the helical bundle and form the greater interaction with the native lipids and SMA polymer and may represent a more dynamic region of the protein. This work shows the promise of using an SMA approach for single particle cryo-EM studies to provide sub-nm structures.",
keywords = "SMALP , Electron microscopy , membrane proteins , AcrB",
author = "Mayuriben Parmar and Shaun Rawson and Scarff, {Charlotte A.} and Adrian Goldman and Dafforn, {Timothy R.} and Muench, {Stephen P.} and Postis, {Vincent L. G.}",
year = "2018",
month = feb,
day = "2",
doi = "10.1016/j.bbamem.2017.10.005",
language = "English",
volume = "1860",
pages = "378--383",
journal = "Biochimica et Biophysica Acta (BBA) - Biomembranes",
issn = "0005-2736",
publisher = "Elsevier",
number = "2",

}

RIS

TY - JOUR

T1 - Using a SMALP platform to determine a sub-nm single particle cryo-EM membrane protein structure

AU - Parmar, Mayuriben

AU - Rawson, Shaun

AU - Scarff, Charlotte A.

AU - Goldman, Adrian

AU - Dafforn, Timothy R.

AU - Muench, Stephen P.

AU - Postis, Vincent L. G.

PY - 2018/2/2

Y1 - 2018/2/2

N2 - The field of membrane protein structural biology has been revolutionized over the last few years with a number of high profile structures being solved using cryo-EM including Piezo, Ryanodine receptor, TRPV1 and the Glutamate receptor. Further developments in the EM field hold the promise of even greater progress in terms of greater resolution, which for membrane proteins is still typically within the 4–7 Å range. One advantage of a cryo-EM approach is the ability to study membrane proteins in more “native” like environments for example proteoliposomes, amphipols and nanodiscs. Recently, styrene maleic acid co-polymers (SMA) have been used to extract membrane proteins surrounded by native lipids (SMALPs) maintaining a more natural environment. We report here the structure of the Escherichia coli multidrug efflux transporter AcrB in a SMALP scaffold to sub-nm resolution, with the resulting map being consistent with high resolution crystal structures and other EM derived maps. However, both the C-terminal helix (TM12) and TM7 are poorly defined in the map. These helices are at the exterior of the helical bundle and form the greater interaction with the native lipids and SMA polymer and may represent a more dynamic region of the protein. This work shows the promise of using an SMA approach for single particle cryo-EM studies to provide sub-nm structures.

AB - The field of membrane protein structural biology has been revolutionized over the last few years with a number of high profile structures being solved using cryo-EM including Piezo, Ryanodine receptor, TRPV1 and the Glutamate receptor. Further developments in the EM field hold the promise of even greater progress in terms of greater resolution, which for membrane proteins is still typically within the 4–7 Å range. One advantage of a cryo-EM approach is the ability to study membrane proteins in more “native” like environments for example proteoliposomes, amphipols and nanodiscs. Recently, styrene maleic acid co-polymers (SMA) have been used to extract membrane proteins surrounded by native lipids (SMALPs) maintaining a more natural environment. We report here the structure of the Escherichia coli multidrug efflux transporter AcrB in a SMALP scaffold to sub-nm resolution, with the resulting map being consistent with high resolution crystal structures and other EM derived maps. However, both the C-terminal helix (TM12) and TM7 are poorly defined in the map. These helices are at the exterior of the helical bundle and form the greater interaction with the native lipids and SMA polymer and may represent a more dynamic region of the protein. This work shows the promise of using an SMA approach for single particle cryo-EM studies to provide sub-nm structures.

KW - SMALP

KW - Electron microscopy

KW - membrane proteins

KW - AcrB

U2 - 10.1016/j.bbamem.2017.10.005

DO - 10.1016/j.bbamem.2017.10.005

M3 - Article

VL - 1860

SP - 378

EP - 383

JO - Biochimica et Biophysica Acta (BBA) - Biomembranes

JF - Biochimica et Biophysica Acta (BBA) - Biomembranes

SN - 0005-2736

IS - 2

ER -