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

Invisible detergents for structure determination of membrane proteins by small‐angle neutron scattering
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通过小角中子散射测定膜蛋白结构的隐形去污剂

DOI:
10.1111/febs.14345
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发表时间:
2018
期刊:
The FEBS Journal
影响因子:
--
通讯作者:
L. Arleth
L. Arleth
中科院分区:
--
文献类型:
--
作者:
S. Midtgaard;T. Darwish;M. C. Pedersen;P. Huda;A. H. Larsen;G. Jensen;S. Kynde;Nicholas Skar;A. Z. Nielsen;Claus Olesen;M. Blaise;J. Dorosz;T. S. Thorsen;R. Venskutonytė;C. Krintel;J. Møller;H. Frielinghaus;E. Gilbert;A. Martel;J. Kastrup;P. Jensen;P. Nissen;L. Arleth

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提出了一种通过小角中子散射(SANS)测定溶液中膜蛋白结构的新方法。用于溶解膜蛋白的常用洗涤剂以同位素取代的形式合成,以利用氢和氘之间的固有中子散射长度差异。实现洗涤剂头部和尾部基团的单独氢/氘水平,使得当通过中子研究时,所形成的胶束在重水(D2 O)中变得有效地不可见。这样,只有来自膜蛋白的信号保留在SANS数据中。我们表明,该方法不仅是普遍适用于五个非常不同的膜蛋白,但也揭示了微妙的结构细节的肌/内质网Ca 2 + ATP酶(SERCA)。总之,同位素取代的洗涤剂的合成使得非专业人员可以通过SANS和随后的数据分析来确定膜蛋白的溶液结构。
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.
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