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

FEBS J. 2018 Jan;285(2):357-371. doi: 10.1111/febs.14345. Epub 2017 Dec 30.

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 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Detergents / chemistry*
  • Glucosides / chemistry*
  • Maltose / analogs & derivatives*
  • Maltose / chemistry
  • Membrane Proteins / chemistry*
  • Micelles
  • Neutron Diffraction*
  • Protein Conformation
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases / chemistry*
  • Scattering, Small Angle*

Substances

  • Detergents
  • Glucosides
  • Membrane Proteins
  • Micelles
  • n-lauryl beta-d-maltopyranoside
  • octyl-beta-D-glucoside
  • Maltose
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases

Associated data

  • PDB/1PY6
  • PDB/1MAL
  • PDB/4RKU
  • PDB/4U2P
  • PDB/1T5S
  • PDB/4UU1
  • PDB/4H1W