Merging In-Solution X-ray and Neutron Scattering Data Allows Fine Structural Analysis of Membrane-Protein Detergent Complexes

Gaëtan Dias Mirandela, Giulia Tamburrino, Miloš T. Ivanović, Felix M. Strnad, Olwyn Byron, Tim Rasmussen, Paul A. Hoskisson, Jochen S. Hub*, Ulrich Zachariae, Frank Gabel, Arnaud Javelle

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)
8 Downloads (Pure)

Abstract

In-solution small-angle X-ray and neutron scattering (SAXS/SANS) have become popular methods to characterize the structure of membrane proteins, solubilized by either detergents or nanodiscs. SANS studies of protein-detergent complexes usually require deuterium-labeled proteins or detergents, which in turn often lead to problems in their expression or purification. Here, we report an approach whose novelty is the combined analysis of SAXS and SANS data from an unlabeled membrane protein complex in solution in two complementary ways. First, an explicit atomic analysis, including both protein and detergent molecules, using the program WAXSiS, which has been adapted to predict SANS data. Second, the use of MONSA which allows one to discriminate between detergent head- and tail-groups in an ab initio approach. Our approach is readily applicable to any detergent-solubilized protein and provides more detailed structural information on protein-detergent complexes from unlabeled samples than SAXS or SANS alone.

Original languageEnglish
Pages (from-to)3910-3914
Number of pages5
JournalJournal of Physical Chemistry Letters
Volume9
Issue number14
Early online date25 Jun 2018
DOIs
Publication statusPublished - 19 Jul 2018

Bibliographical note

G.D.M. and A.J. were supported by a Ph.D. and a Chancellor’s
Fellowship from Strathclyde University, respectively, G.T. and
U.Z. acknowledge funding from the Scottish Universities’
Physics Alliance (SUPA). P.A.H. acknowledges the support of
the Natural Environment Research Council (NE/M001415/1)
and A.J. the support of Tenovus Scotland (Project S17-07).
M.T.I., F.M.S., and J.S.H. acknowledge support by the Deutsche
Forschungsgemeinschaft (HU 1971-1/1, HU 1971-3/1, HU
1971-4/1). We thank the ILL Block Allocation Group (BAG)
system for SANS beamtime at D22 and Dr A. Martel for help
with the setup of the instrument. We acknowledge Diamond
Light Source for time on Beamline B21. We thanks Dr. P. Soule
(NanoTemper Technologies GmbH) and Dr. M. Tully
(DIAMOND, U.K.) for help with the microscale thermophoresis
experiments and SEC-SAXS data acquisition, respectively

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