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

Selma Maric; Nicholas Skar-Gislinge; Søren Midtgaard; Mikkel Boas Thygesen; Jürgen Schiller; Henrich Frielinghaus; Martine Moulin; Michael Haertlein; V. Trevor Forsyth; Thomas Günther-Pomorski; Lise Arleth

doi:10.1107/s1399004713027466

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

Selma Maric, Nicholas Skar-Gislinge, Søren Midtgaard, Mikkel Boas Thygesen, Jürgen Schiller, Henrich Frielinghaus, Martine Moulin, Michael Haertlein, V. Trevor Forsyth, Thomas Günther-Pomorski, Lise Arleth^*

^*Corresponding author for this work

38 Citations (Scopus)

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.

Original language	English
Journal	Acta Crystallographica. Section D: Biological Crystallography
Volume	70
Issue number	2
Pages (from-to)	317-328
Number of pages	12
ISSN	0907-4449
DOIs	https://doi.org/10.1107/s1399004713027466
Publication status	Published - 2014

Keywords

contrast variation
phospholipid bilayer nanodisc
small-angle neutron scattering

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Maric, S., Skar-Gislinge, N., Midtgaard, S., Thygesen, M. B., Schiller, J., Frielinghaus, H., Moulin, M., Haertlein, M., Forsyth, V. T., Günther-Pomorski, T., & Arleth, L. (2014). Stealth carriers for low-resolution structure determination of membrane proteins in solution. Acta Crystallographica. Section D: Biological Crystallography, 70(2), 317-328. https://doi.org/10.1107/s1399004713027466

Maric, S, Skar-Gislinge, N, Midtgaard, S, Thygesen, MB, Schiller, J, Frielinghaus, H, Moulin, M, Haertlein, M, Forsyth, VT, Günther-Pomorski, T & Arleth, L 2014, 'Stealth carriers for low-resolution structure determination of membrane proteins in solution', Acta Crystallographica. Section D: Biological Crystallography, vol. 70, no. 2, pp. 317-328. https://doi.org/10.1107/s1399004713027466

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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.",

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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

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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

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DO - 10.1107/s1399004713027466

M3 - Journal article

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AN - SCOPUS:84894134926

SN - 0907-4449

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EP - 328

JO - Acta Crystallographica. Section D: Biological Crystallography

JF - Acta Crystallographica. Section D: Biological Crystallography

IS - 2

ER -

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

Abstract

Keywords

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