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

Midtgaard, Søren Roi ; Darwish, Tamim A ; Pedersen, Martin Cramer ; Huda, Pie ; Larsen, Andreas Haahr ; Jensen, Grethe Vestergaard ; Kynde, Søren Andreas Røssel ; Skar-Gislinge, Nicholas ; Nielsen, Agnieszka Janina Zygadlo and Olesen, Claus , et al. (2018) In The FEBS Journal 285(2). p.357-371
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 (D2 O) 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... (More)

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 (D2 O) 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 non-specialists. This article is protected by copyright. All rights reserved.

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@article{35ac359d-bd09-4b43-a41b-af57403f3534,
  abstract     = {{<p>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 (D2 O) 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 non-specialists. This article is protected by copyright. All rights reserved.</p>}},
  author       = {{Midtgaard, Søren Roi and Darwish, Tamim A and Pedersen, Martin Cramer and Huda, Pie and Larsen, Andreas Haahr and Jensen, Grethe Vestergaard and Kynde, Søren Andreas Røssel and Skar-Gislinge, Nicholas and Nielsen, Agnieszka Janina Zygadlo and Olesen, Claus and Blaise, Mickael and Dorosz, Jerzy Józef and Thorsen, Thor Seneca and Venskutonytė, Raminta and Krintel, Christian and Møller, Jesper V and Frielinghaus, Henrich and Gilbert, Elliot Paul and Martel, Anne and Kastrup, Jette Sandholm and Jensen, Poul Erik and Nissen, Poul and Arleth, Lise}},
  issn         = {{1742-464X}},
  language     = {{eng}},
  number       = {{2}},
  pages        = {{357--371}},
  publisher    = {{Wiley-Blackwell}},
  series       = {{The FEBS Journal}},
  title        = {{Invisible detergents for structure determination of membrane proteins by small-angle neutron scattering}},
  url          = {{http://dx.doi.org/10.1111/febs.14345}},
  doi          = {{10.1111/febs.14345}},
  volume       = {{285}},
  year         = {{2018}},
}