Acta Crystallographica Section D

Biological Crystallography

Volume 70, Part 2 (February 2014)

research papers

Acta Cryst. (2014). D70, 317-328    [ doi:10.1107/S1399004713027466 ]

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

S. Maric, N. Skar-Gislinge, S. Midtgaard, M. B. Thygesen, J. Schiller, H. Frielinghaus, M. Moulin, M. Haertlein, V. T. Forsyth, T. G. Pomorski and L. Arleth

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 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.

Keywords: phospholipid bilayer nanodisc; small-angle neutron scattering; contrast variation.

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[ doi:10.1107/S1399004713027466/kw5078sup1.pdf ]
Assembly of deuterated stealth nanodiscs.


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