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Conformational States of ABC Transporter MsbA in a Lipid Environment Investigated by Small-Angle Scattering Using Stealth Carrier Nanodiscs

Josts, Inokentijs ; Nitsche, Julius ; Maric, Selma LU ; Mertens, Haydyn D ; Moulin, Martine ; Haertlein, Michael ; Prevost, Sylvain ; Svergun, Dmitri I ; Busch, Sebastian and Forsyth, V Trevor , et al. (2018) In Structure 26(8). p.4-1079
Abstract

Structural studies of integral membrane proteins (IMPs) are challenging, as many of them are inactive or insoluble in the absence of a lipid environment. Here, we describe an approach making use of fractionally deuterium labeled "stealth carrier" nanodiscs that are effectively invisible to low-resolution neutron diffraction and enable structural studies of IMPs in a lipidic native-like solution environment. We illustrate the potential of the method in a joint small-angle neutron scattering (SANS) and X-ray scattering (SAXS) study of the ATP-binding cassette (ABC) transporter protein MsbA solubilized in the stealth nanodiscs. The data allow for a direct observation of the signal from the solubilized protein without contribution from the... (More)

Structural studies of integral membrane proteins (IMPs) are challenging, as many of them are inactive or insoluble in the absence of a lipid environment. Here, we describe an approach making use of fractionally deuterium labeled "stealth carrier" nanodiscs that are effectively invisible to low-resolution neutron diffraction and enable structural studies of IMPs in a lipidic native-like solution environment. We illustrate the potential of the method in a joint small-angle neutron scattering (SANS) and X-ray scattering (SAXS) study of the ATP-binding cassette (ABC) transporter protein MsbA solubilized in the stealth nanodiscs. The data allow for a direct observation of the signal from the solubilized protein without contribution from the surrounding lipid nanodisc. Not only the overall shape but also differences between conformational states of MsbA can be reliably detected from the scattering data, demonstrating the sensitivity of the approach and its general applicability to structural studies of IMPs.

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publishing date
type
Contribution to journal
publication status
published
in
Structure
volume
26
issue
8
pages
4 - 1079
publisher
Cell Press
external identifiers
  • pmid:29937358
  • scopus:85047450164
ISSN
0969-2126
DOI
10.1016/j.str.2018.05.007
language
English
LU publication?
no
id
d5289311-4f71-4d34-aac3-db553262ef42
date added to LUP
2018-07-19 12:05:22
date last changed
2024-06-11 17:55:03
@article{d5289311-4f71-4d34-aac3-db553262ef42,
  abstract     = {{<p>Structural studies of integral membrane proteins (IMPs) are challenging, as many of them are inactive or insoluble in the absence of a lipid environment. Here, we describe an approach making use of fractionally deuterium labeled "stealth carrier" nanodiscs that are effectively invisible to low-resolution neutron diffraction and enable structural studies of IMPs in a lipidic native-like solution environment. We illustrate the potential of the method in a joint small-angle neutron scattering (SANS) and X-ray scattering (SAXS) study of the ATP-binding cassette (ABC) transporter protein MsbA solubilized in the stealth nanodiscs. The data allow for a direct observation of the signal from the solubilized protein without contribution from the surrounding lipid nanodisc. Not only the overall shape but also differences between conformational states of MsbA can be reliably detected from the scattering data, demonstrating the sensitivity of the approach and its general applicability to structural studies of IMPs.</p>}},
  author       = {{Josts, Inokentijs and Nitsche, Julius and Maric, Selma and Mertens, Haydyn D and Moulin, Martine and Haertlein, Michael and Prevost, Sylvain and Svergun, Dmitri I and Busch, Sebastian and Forsyth, V Trevor and Tidow, Henning}},
  issn         = {{0969-2126}},
  language     = {{eng}},
  month        = {{05}},
  number       = {{8}},
  pages        = {{4--1079}},
  publisher    = {{Cell Press}},
  series       = {{Structure}},
  title        = {{Conformational States of ABC Transporter MsbA in a Lipid Environment Investigated by Small-Angle Scattering Using Stealth Carrier Nanodiscs}},
  url          = {{http://dx.doi.org/10.1016/j.str.2018.05.007}},
  doi          = {{10.1016/j.str.2018.05.007}},
  volume       = {{26}},
  year         = {{2018}},
}