Invisible detergents for structure determination of membrane proteins by small-angle neutron scattering

Søren Roi Midtgaard; Tamim A. Darwish; Martin Cramer Pedersen; Pie Huda; Andreas Haahr Larsen; Grethe Vestergaard Jensen; Søren Andreas Røssell Kynde; Nicholas Skar-Gislinge; Agnieszka Janina Zygadlo Nielsen; Claus Olesen; Mickael Blaise; Jerzy Józef Dorosz; Thor Seneca Thorsen; Raminta Venskutonytė; Christian Krintel; Jesper V. Møller; Henrich Frielinghaus; Elliot Paul Gilbert; Anne Martel; Jette Sandholm Kastrup; Poul Erik Jensen; Poul Nissen; Lise Arleth

doi:10.1111/febs.14345

Invisible detergents for structure determination of membrane proteins by small-angle neutron scattering

Søren Roi Midtgaard^*, Tamim A. Darwish, Martin Cramer Pedersen, Pie Huda, Andreas Haahr Larsen, Grethe Vestergaard Jensen, Søren Andreas Røssell Kynde, Nicholas Skar-Gislinge, Agnieszka Janina Zygadlo Nielsen, Claus Olesen, Mickael Blaise, Jerzy Józef Dorosz, Thor Seneca Thorsen, Raminta Venskutonytė, Christian Krintel, Jesper V. Møller, Henrich Frielinghaus, Elliot Paul Gilbert, Anne Martel, Jette Sandholm KastrupPoul Erik Jensen, Poul Nissen, Lise Arleth

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

41 Citations (Scopus)

Abstract

A novel and generally applicable method for determining structures of membrane proteins in solution via small-angle neutron scattering (SANS) is presented. Common detergents for solubilizing membrane proteins were synthesized in isotope-substituted versions for utilizing the intrinsic neutron scattering length difference between hydrogen and deuterium. Individual hydrogen/deuterium levels of the detergent head and tail groups were achieved such that the formed micelles became effectively invisible in heavy water (D₂O) when investigated by neutrons. This way, only the signal from the membrane protein remained in the SANS data. We demonstrate that the method is not only generally applicable on five very different membrane proteins but also reveals subtle structural details about the sarco/endoplasmatic reticulum Ca²⁺ ATPase (SERCA). In all, the synthesis of isotope-substituted detergents makes solution structure determination of membrane proteins by SANS and subsequent data analysis available to nonspecialists.

Original language	English
Pages (from-to)	357-371
Number of pages	15
Journal	FEBS Journal
Volume	285
Issue number	2
DOIs	https://doi.org/10.1111/febs.14345
Publication status	Published - Jan 2018

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Midtgaard, S. R., Darwish, T. A., Pedersen, M. C., Huda, P., Larsen, A. H., Jensen, G. V., Kynde, S. A. R., Skar-Gislinge, N., Nielsen, A. J. Z., Olesen, C., Blaise, M., Dorosz, J. J., Thorsen, T. S., Venskutonytė, R., Krintel, C., Møller, J. V., Frielinghaus, H., Gilbert, E. P., Martel, A., ... Arleth, L. (2018). Invisible detergents for structure determination of membrane proteins by small-angle neutron scattering. FEBS Journal, 285(2), 357-371. https://doi.org/10.1111/febs.14345

@article{583a87f6baff4400b0fd8f7d4da680e9,

title = "Invisible detergents for structure determination of membrane proteins by small-angle neutron scattering",

abstract = "A novel and generally applicable method for determining structures of membrane proteins in solution via small-angle neutron scattering (SANS) is presented. Common detergents for solubilizing membrane proteins were synthesized in isotope-substituted versions for utilizing the intrinsic neutron scattering length difference between hydrogen and deuterium. Individual hydrogen/deuterium levels of the detergent head and tail groups were achieved such that the formed micelles became effectively invisible in heavy water (D2O) when investigated by neutrons. This way, only the signal from the membrane protein remained in the SANS data. We demonstrate that the method is not only generally applicable on five very different membrane proteins but also reveals subtle structural details about the sarco/endoplasmatic reticulum Ca2+ ATPase (SERCA). In all, the synthesis of isotope-substituted detergents makes solution structure determination of membrane proteins by SANS and subsequent data analysis available to nonspecialists.",

keywords = "SANS, Small-angle neutron scattering, contrast matching, deuteration, membrane proteins",

author = "Midtgaard, {S{\o}ren Roi} and Darwish, {Tamim A.} and Pedersen, {Martin Cramer} and Pie Huda and Larsen, {Andreas Haahr} and Jensen, {Grethe Vestergaard} and Kynde, {S{\o}ren Andreas R{\o}ssell} and Nicholas Skar-Gislinge and Nielsen, {Agnieszka Janina Zygadlo} and Claus Olesen and Mickael Blaise and Dorosz, {Jerzy J{\'o}zef} and Thorsen, {Thor Seneca} and Raminta Venskutonytė and Christian Krintel and M{\o}ller, {Jesper V.} and Henrich Frielinghaus and Gilbert, {Elliot Paul} and Anne Martel and Kastrup, {Jette Sandholm} and Jensen, {Poul Erik} and Poul Nissen and Lise Arleth",

note = "Publisher Copyright: {\textcopyright} 2017 Federation of European Biochemical Societies",

year = "2018",

month = jan,

doi = "10.1111/febs.14345",

language = "English",

volume = "285",

pages = "357--371",

journal = "FEBS Journal",

issn = "1742-464X",

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Midtgaard, SR, Darwish, TA, Pedersen, MC, Huda, P, Larsen, AH, Jensen, GV, Kynde, SAR, Skar-Gislinge, N, Nielsen, AJZ, Olesen, C, Blaise, M, Dorosz, JJ, Thorsen, TS, Venskutonytė, R, Krintel, C, Møller, JV, Frielinghaus, H, Gilbert, EP, Martel, A, Kastrup, JS, Jensen, PE, Nissen, P & Arleth, L 2018, 'Invisible detergents for structure determination of membrane proteins by small-angle neutron scattering', FEBS Journal, vol. 285, no. 2, pp. 357-371. https://doi.org/10.1111/febs.14345

TY - JOUR

T1 - Invisible detergents for structure determination of membrane proteins by small-angle neutron scattering

AU - Midtgaard, Søren Roi

AU - Darwish, Tamim A.

AU - Pedersen, Martin Cramer

AU - Huda, Pie

AU - Larsen, Andreas Haahr

AU - Jensen, Grethe Vestergaard

AU - Kynde, Søren Andreas Røssell

AU - Skar-Gislinge, Nicholas

AU - Nielsen, Agnieszka Janina Zygadlo

AU - Olesen, Claus

AU - Blaise, Mickael

AU - Dorosz, Jerzy Józef

AU - Thorsen, Thor Seneca

AU - Venskutonytė, Raminta

AU - Krintel, Christian

AU - Møller, Jesper V.

AU - Frielinghaus, Henrich

AU - Gilbert, Elliot Paul

AU - Martel, Anne

AU - Kastrup, Jette Sandholm

AU - Jensen, Poul Erik

AU - Nissen, Poul

AU - Arleth, Lise

PY - 2018/1

Y1 - 2018/1

N2 - A novel and generally applicable method for determining structures of membrane proteins in solution via small-angle neutron scattering (SANS) is presented. Common detergents for solubilizing membrane proteins were synthesized in isotope-substituted versions for utilizing the intrinsic neutron scattering length difference between hydrogen and deuterium. Individual hydrogen/deuterium levels of the detergent head and tail groups were achieved such that the formed micelles became effectively invisible in heavy water (D2O) when investigated by neutrons. This way, only the signal from the membrane protein remained in the SANS data. We demonstrate that the method is not only generally applicable on five very different membrane proteins but also reveals subtle structural details about the sarco/endoplasmatic reticulum Ca2+ ATPase (SERCA). In all, the synthesis of isotope-substituted detergents makes solution structure determination of membrane proteins by SANS and subsequent data analysis available to nonspecialists.

AB - A novel and generally applicable method for determining structures of membrane proteins in solution via small-angle neutron scattering (SANS) is presented. Common detergents for solubilizing membrane proteins were synthesized in isotope-substituted versions for utilizing the intrinsic neutron scattering length difference between hydrogen and deuterium. Individual hydrogen/deuterium levels of the detergent head and tail groups were achieved such that the formed micelles became effectively invisible in heavy water (D2O) when investigated by neutrons. This way, only the signal from the membrane protein remained in the SANS data. We demonstrate that the method is not only generally applicable on five very different membrane proteins but also reveals subtle structural details about the sarco/endoplasmatic reticulum Ca2+ ATPase (SERCA). In all, the synthesis of isotope-substituted detergents makes solution structure determination of membrane proteins by SANS and subsequent data analysis available to nonspecialists.

KW - SANS

KW - Small-angle neutron scattering

KW - contrast matching

KW - deuteration

KW - membrane proteins

UR - http://www.scopus.com/inward/record.url?scp=85039723710&partnerID=8YFLogxK

U2 - 10.1111/febs.14345

DO - 10.1111/febs.14345

M3 - Article

SN - 1742-464X

VL - 285

SP - 357

EP - 371

JO - FEBS Journal

JF - FEBS Journal

IS - 2

ER -

Invisible detergents for structure determination of membrane proteins by small-angle neutron scattering

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