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Stealth carriers for low-resolution structure determination of membrane proteins in solution

Maric, Selma LU ; Skar-Gislinge, Nicholas ; Midtgaard, Søren ; Thygesen, Mikkel B ; Schiller, Jürgen ; Frielinghaus, Henrich ; Moulin, Martine ; Haertlein, Michael ; Forsyth, V Trevor and Pomorski, Thomas Günther , et al. (2014) In Acta Crystallographica. Section D: Biological Crystallography 70(Pt 2). p.28-317
Abstract

Structural studies of membrane proteins remain a great experimental challenge. Functional reconstitution into artificial nanoscale bilayer disc carriers that mimic the native bilayer environment allows the handling of membrane proteins in solution. This enables the use of small-angle scattering techniques for fast and reliable structural analysis. The difficulty with this approach is that the carrier discs contribute to the measured scattering intensity in a highly nontrivial fashion, making subsequent data analysis challenging. Here, an elegant solution to circumvent the intrinsic complexity brought about by the presence of the carrier disc is presented. In combination with small-angle neutron scattering (SANS) and the D2O/H2O-based... (More)

Structural studies of membrane proteins remain a great experimental challenge. Functional reconstitution into artificial nanoscale bilayer disc carriers that mimic the native bilayer environment allows the handling of membrane proteins in solution. This enables the use of small-angle scattering techniques for fast and reliable structural analysis. The difficulty with this approach is that the carrier discs contribute to the measured scattering intensity in a highly nontrivial fashion, making subsequent data analysis challenging. Here, an elegant solution to circumvent the intrinsic complexity brought about by the presence of the carrier disc is presented. In combination with small-angle neutron scattering (SANS) and the D2O/H2O-based solvent contrast-variation method, it is demonstrated that it is possible to prepare specifically deuterated carriers that become invisible to neutrons in 100% D2O at the length scales relevant to SANS. These `stealth' carrier discs may be used as a general platform for low-resolution structural studies of membrane proteins using well established data-analysis tools originally developed for soluble proteins.

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publication status
published
keywords
Deuterium/chemistry, Escherichia coli/genetics, Lipid Bilayers/chemistry, Membrane Proteins/chemistry, Membranes, Artificial, Models, Molecular, Neutron Diffraction, Neutrons, Phosphatidylcholines/chemistry, Protein Conformation, Recombinant Proteins/chemistry, Scattering, Small Angle
in
Acta Crystallographica. Section D: Biological Crystallography
volume
70
issue
Pt 2
pages
12 pages
publisher
John Wiley and Sons Inc.
external identifiers
  • pmid:24531466
  • scopus:84894134926
ISSN
1399-0047
DOI
10.1107/S1399004713027466
language
English
LU publication?
no
id
61d5c184-1e03-4d03-88de-aa227622863b
date added to LUP
2018-07-19 11:57:27
date last changed
2020-08-12 08:07:44
@article{61d5c184-1e03-4d03-88de-aa227622863b,
  abstract     = {<p>Structural studies of membrane proteins remain a great experimental challenge. Functional reconstitution into artificial nanoscale bilayer disc carriers that mimic the native bilayer environment allows the handling of membrane proteins in solution. This enables the use of small-angle scattering techniques for fast and reliable structural analysis. The difficulty with this approach is that the carrier discs contribute to the measured scattering intensity in a highly nontrivial fashion, making subsequent data analysis challenging. Here, an elegant solution to circumvent the intrinsic complexity brought about by the presence of the carrier disc is presented. In combination with small-angle neutron scattering (SANS) and the D2O/H2O-based solvent contrast-variation method, it is demonstrated that it is possible to prepare specifically deuterated carriers that become invisible to neutrons in 100% D2O at the length scales relevant to SANS. These `stealth' carrier discs may be used as a general platform for low-resolution structural studies of membrane proteins using well established data-analysis tools originally developed for soluble proteins. </p>},
  author       = {Maric, Selma and Skar-Gislinge, Nicholas and Midtgaard, Søren and Thygesen, Mikkel B and Schiller, Jürgen and Frielinghaus, Henrich and Moulin, Martine and Haertlein, Michael and Forsyth, V Trevor and Pomorski, Thomas Günther and Arleth, Lise},
  issn         = {1399-0047},
  language     = {eng},
  number       = {Pt 2},
  pages        = {28--317},
  publisher    = {John Wiley and Sons Inc.},
  series       = {Acta Crystallographica. Section D: Biological Crystallography},
  title        = {Stealth carriers for low-resolution structure determination of membrane proteins in solution},
  url          = {http://dx.doi.org/10.1107/S1399004713027466},
  doi          = {10.1107/S1399004713027466},
  volume       = {70},
  year         = {2014},
}