TY - JOUR
T1 - Stealth carriers for low-resolution structure determination of membrane proteins in solution
AU - Maric, Selma
AU - Skar-Gislinge, Nicholas
AU - Midtgaard, Søren
AU - Thygesen, Mikkel Boas
AU - Schiller, Jürgen
AU - Frielinghaus, Henrich
AU - Moulin, Martine
AU - Haertlein, Michael
AU - Forsyth, V. Trevor
AU - Günther-Pomorski, Thomas
AU - Arleth, Lise
PY - 2014
Y1 - 2014
N2 - 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.
AB - 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.
KW - contrast variation
KW - phospholipid bilayer nanodisc
KW - small-angle neutron scattering
U2 - 10.1107/s1399004713027466
DO - 10.1107/s1399004713027466
M3 - Journal article
C2 - 24531466
AN - SCOPUS:84894134926
SN - 0907-4449
VL - 70
SP - 317
EP - 328
JO - Acta Crystallographica. Section D: Biological Crystallography
JF - Acta Crystallographica. Section D: Biological Crystallography
IS - 2
ER -